BrdU Antibody | BU1/75 (ICR1)

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Formalin fixed, paraffin embedded mouse colon stained with Rat anti BrdU (MCA2060): low power

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Formalin fixed, paraffin embedded mouse colon stained with Rat anti BrdU (MCA2060): high power

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Formalin fixed, paraffin embedded mouse colon stained with Rat anti BrdU (MCA2060): med power

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HL-60 cells were pulse labeled with BrdU for 45 minutes prior to harvesting and then incubated with primary antibody Rat anti BrdU (MCA2060GA) clone BU1/75 diluted 1/100 followed by Rabbit anti Rat IgG secondary antibody (STAR17B) FITC-conjugated, diluted 1/200

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Rat anti BrdU antibody, clone Bu1/75 used for the detection of proliferating cells in mouse brain by immunohistochemistry on free-floating sections.
Image caption:
Impaired base-line proliferation in the hippocampus in the absence of αβ T cells. ( A) Representative BrdU immunohistochemistry of the hippocampal dentate gyrus from eight week-old wild-type (A1), TCRα +/- (A2) and TCRα -/- (A3) mice, 24 hours after the first of 3 consecutive BrdU injections. Scale bar, 100 μm. ( B) Quantification of BrdU + cells in the dentate gyrus of wild-type (n = 6), TCRα +/- (n = 7) and TCRα -/- (n = 8) mice. All numbers are mean ± SEM. ANOVA, * p < 0.05.

From: Niebling J, E. Rünker A, Schallenberg S et al. Myelin-specific T helper 17 cells promote adult hippocampal neurogenesis through indirect mechanisms
F1000Research 2014, 3:169.

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Rat anti BrdU antibody, clone Bu1/75 used for the detection of BrdU positive cells in mouse brain by immunofluorescence.
Image caption:
Disruption of the Stx1 function impairs GBM tumor progression in vivo. An equal number of the indicated U373 cell populations were stereotactically inoculated into the brain of athymic nude mice. The size of the tumors was estimated at different days post-inoculation by the quantification of luciferase activity in the tumor cells. A, D. Representative images of the luciferase signal from mice inoculated with the indicated U373 GBM cells after 40 DPI. B, E. Growth curves of the indicated U373 GBM tumors showing a marked reduction of the tumor sizes after impairment of Stx1 function (* p ≤ 0.05 at 40 DPI). C, F. Scatter plots showing the individual size of the indicated U373 GBM tumors after 40 DPI. G. Representative entire brain NMR images of mice inoculated with the indicated U373 cell populations after 30 DPI (arrowheads indicate the tumors formed) showing a marked reduction in the Stx1a-Sh01 and Stx1-DN cells. H. Scatter plot showing the area of the indicated U373 GBM tumors formed after 30 DPI. I. Representative confocal images from histological sections of 30 DPI brain tumors stained with anti-GFP and anti-BrdU antibodies. J. Quantification of BrdU-positive nuclei in 30 DPI U373 cell tumors showing that Stx1 loss-of-function reduces proliferation (* p ≤ 0.05; *** p ≤ 0.001).

From: Ulloa F, Gonzàlez-Juncà A, Meffre D, Barrecheguren PJ, Martínez-Mármol R, et al. (2015) Blockade of the SNARE Protein Syntaxin 1 Inhibits Glioblastoma Tumor Growth.
PLoS ONE 10(3): e0119707.

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Rat anti BrdU antibody, clone Bu1/75 used for the detection of BrdU incorporating cells in mouse colon by immunofluorescence.
Image caption:
VIP enhances colonic crypt function at baseline. Exogenous VIP was administered to VIPKO mice (VIPKO-VIP) daily for 10 days. Immunostaining for Ki67 and quantitative analysis of Ki67+ve cells (A). Immunostaining for BrdU+ve cells and calculated spatial distribution at 72h post injection (B). Quantitative analysis of TUNEL +ve crypt IEC (C). Epithelial permeability measured by FITC dextran (D); n = 6–10 animals/group, results are represented as means ± SEM, *P< 0.05, **P< 0.01, ***P< 0.001. Scale bar = 50 μm.

From: Wu X, Conlin VS, Morampudi V, Ryz NR, Nasser Y, Bhinder G, et al. (2015) Vasoactive Intestinal Polypeptide Promotes Intestinal Barrier Homeostasis and Protection Against Colitis in Mice.
PLoS ONE 10(5): e0125225.

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Rat anti BrdU antibody, clone Bu1/75 used for the detection of BrdU incorporating cells in the intestine of rats by immunohistochemistry on formalin fixed, paraffin embedded tissue sections.
Image caption:
BrdU labeling of the small intestine after i.v. administration. BrdU (50 mg/kg b.wt.), a thymidine analogue, was given 2 h prior to removing the intestine. The dark labeling by BrdU is exclusively located to the nuclei of intestinal crypt cells. Mayer's haematoxylin.

From: Lundgren O, Jodal M, Jansson M, Ryberg AT, Svensson L (2011)
Intestinal Epithelial Stem/Progenitor Cells Are Controlled by Mucosal Afferent Nerves.
PLoS ONE 6(2): e16295.

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Rat anti BrdU antibody, clone Bu1/75 used for the detection of dividing cells in mouse embryo by immunofluorescence on free floating, formalin fixed paraffin embedded tissue sections.
Image caption:
Proliferative potential of EGFP-labeled cells. One day or 14 days after the last TAM injection, dividing cells were labeled via a single BrdU injection and co-localization of EGFP with BrdU was examined at day 2 (a and c) and day 15 (b and d), respectively, in the SVZ and DG. Quantification on day 2 revealed that the percentage of all EGFP+ cells labeled with BrdU rapidly decreases and is shown in (e). Scale bar in (c) = 33 μm.

From: Zhang J, Giesert F, Kloos K, Vogt Weisenhorn DM, Aigner L, Wurst W, Couillard-Despres S. A powerful transgenic tool for fate mapping and functional analysis of newly generated neurons.
BMC Neurosci. 2010 Dec 31;11:158.

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Rat anti BrdU antibody, clone Bu1/75 used for the detection of proliferating cells in the rat kidney by immunohistochemistry on formalin fixed, paraffing embedded tissue sections.
Image caption:
Presence of BrdU-positive cells in the normal rat kidneys after long-term BrdU labeling. a Experimental design. BrdU was intraperitoneally injected into normal rats for the indicated periods. After BrdU labeling, the rats were sacrificed, and the kidneys were removed and used for histological examination. In another experiment, UUO was induced in rats pre-treated with BrdU for 4 weeks as described in “Methods”. At the indicated times after UUO, the rats were sacrificed and the kidneys were removed and used for histological examination. b, c Detection of BrdU-positive cells in the normal rat kidneys after long-term BrdU labeling. BrdU was injected intraperitoneally into the normal rats for the indicated periods. BrdU-positive cells (brown nuclei) were examined by immunostaining and were counterstained with PAS. Magnification is ×400 in b and ×1000 in c. CO cortex, OM outer medulla, IM inner medulla. d Quantitative analysis of BrdU-positive cells. (Filled square) OM, outer medulla, (empty circle) CO, cortex, (filled triangle) IM, inner medulla. Values are means?±?SE (n?=?5).

From: Nakasatomi M, Maeshima A, Mishima K, Ikeuchi H, Sakairi T, Kaneko Y, Hiromura K, Nojima Y. Novel approach for the detection of tubular cell migration into the interstitium during renal fibrosis in rats.
Fibrogenesis Tissue Repair. 2015 Jul 10;8:12.

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Rat anti BrdU antibody used for the identification of proliferating cells in the axolotl brain by immunohistochemistry on formalin fixed, paraffin embedded tissue sections.
Image caption:
Cells labeled with BrdU after an overnight incubation are largely retained in the ventricular zone throughout the neuraxis, which shows regional variation in proliferative activity. (A-F) Examples of BrdU+ cells in the VZ of different brain regions show variation in numbers of labeled cells. Regions of the brain are marked. In D and F, BrdU+ cells are marked with red arrowheads. (G) Counts of proliferating VZ cells in different regions of the brain expressed as a percentage of total VZ cells. at: anterior telencephalon; BrdU: bromodeoxyuridine; cereb: cerebellum; dlt: dorsolateral telencephalon; mes: mesencephalon; mlt: mediolateral telencephalon; rh: rhombencephalon; vt: ventral telencephalon; VZ: ventricular zone. * significant (P = 0.025), ** very significant (P = 0.006), *** extremely significant (P = 0.0004) by Student’s T-test.

From: Maden M, Manwell LA, Ormerod BK. Proliferation zones in the axolotl brain and regeneration of the telencephalon.
Neural Dev. 2013 Jan 17;8:1.

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Rat anti BrdU antibody used for the identification of proliferating cells in the axolotl brain by immunohistochemistry on formalin fixed, paraffin embedded tissue sections.
Image caption:
Sections taken one week after overnight BrdU labeling show the migration of BrdU+ cells from the ventricular zone into the mantle zone at different rates at different levels of the central nervous system. (A, C, F, G, I, J) Positions of labeled cells on five cumulative sections at the levels marked on the figure. The migration of BrdU+ cells outwards at different rates can be appreciated by comparing A and C with G. (B, D, E, H) Labeled cells on individual sections.

From: Maden M, Manwell LA, Ormerod BK. Proliferation zones in the axolotl brain and regeneration of the telencephalon.
Neural Dev. 2013 Jan 17;8:1.

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Rat anti BrdU antibody used for the identification of proliferating cells in the axolotl brain by immunohistochemistry on formalin fixed, paraffin embedded tissue sections.
Image caption:
Migration patterns and glial fibrillary acidic protein expression in the telencephalon. (A) Single section through the anterior telencephalon and olfactory bulb taken 4 weeks after overnight BrdU labeling shows a stream of cells moving laterally into the olfactory bulb. (B) Cumulative positions of BrdU+ cells in the anterior telencephalon and olfactory bulb level 4 weeks after overnight labeling. (C) Cumulative positions of BrdU+ cells in the anterior telencephalon and olfactory nerve level 4 weeks after overnight labeling showing a decreased number of BrdU+ cells in the VZ. (D) Cumulative positions of BrdU+ cells in the posterior telencephalon 4 weeks after overnight labeling showing a decreased number of BrdU+ cells in the VZ. (E,F) Patterns of BrdU+ cells in and adjacent to the VZ 3 weeks after overnight labeling. (G,H) Glial fibrillary acidic protein immunocytochemistry of the normal telencephalon at low power (G) and high power (H) to show the cytoplasmic extensions of radial glial cells. BrdU: bromodeoxyuridine; VZ: ventricular zone.

From: Maden M, Manwell LA, Ormerod BK. Proliferation zones in the axolotl brain and regeneration of the telencephalon.
Neural Dev. 2013 Jan 17;8:1.

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Rat anti BrdU antibody used for the identification of proliferating cells in the axolotl brain by immunohistochemistry on formalin fixed, paraffin embedded tissue sections.
Image caption:
Ventricular zones with variable proliferative activity can be detected throughout the axolotl central nervous system. (A) Structure of the axolotl brain showing the levels at which sections ventricular zone proliferation was analyzed. B = anterior telencephalon and olfactory nerve, C = anterior telencephalon and olfactory bulb, D = posterior telencephalon, E = mesencephalon, F = cerebellum, G = rhombencephalon; on: olfactory nerve. Scale bar = 1?mm. (B – G) Cumulative BrdU+ cell numbers on five sections from each level depicted in A on which individual labeled cells are marked with a red dot. Regions of the brain are marked.; on: olfactory nerve; ob: olfactory bulb; cp: choroid plexus.

From: Maden M, Manwell LA, Ormerod BK. Proliferation zones in the axolotl brain and regeneration of the telencephalon.
Neural Dev. 2013 Jan 17;8:1.

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Rat anti BrdU antibody used for the identification of proliferating cells in the axolotl brain by immunohistochemistry on formalin fixed, paraffin embedded tissue sections.
Image caption:
Two weeks after BrdU labeling, new cells have migrated and differentiated into neurons and a few new cells are retained in the ventricular zone and express GFAP. (A) Confocal image at 10× magnification shows that, in the uninjured axolotl telencephalon, most BrdU+ cells have migrated into the mantle zone 2 weeks after labeling. A few BrdU+ cells are retained in the VZ. (B,C) Confocal images taken at 40× objective (2.3× digital zoom) of sections stained with 4'-6-diamidino-2-phenylindole (in white; B1 and C1) and antibodies against (B) BrdU (in red; B2) and GFAP (in green; B3) or (C) BrdU (in red; C2), DCX (in blue; C3), NeuN (in green; C3). The image in (B) shows that very few BrdU+ cells are retained in the VZ by 2 weeks after overnight BrdU labeling, but those that do remain and express GFAP may represent new radial glia-like cells. (C) by 2 weeks, all BrdU+ cells in the mantle zone express the mature neuronal marker NeuN (white and yellow arrows) and a small percentage of the NeuN+ neurons retain DCX expression (yellow arrows) and are likely transitioning into mature neurons. (D) The relatively small densities (versus F) of BrdU/GFAP+ cells retained in the VZ are resilient across weeks 2 to 4 after BrdU labeling. (E) Most BrdU+ VZ cells express GFAP. (F) All BrdU+ cells in the neuronal layer express NeuN and a few NeuN+ neurons retain DCX expression. These percentages are consistent across weeks 2 to 4 after BrdU labeling. (G) New neurons are either vulnerable to cell death or are still migrating because their densities significantly decrease between weeks 2 and 3 (**P <0.001). BrdU: bromodeoxyuridine; DCX: doublecortin; GFAP: glial fibrillary acidic protein; VZ: ventricular zone.

From: Maden M, Manwell LA, Ormerod BK. Proliferation zones in the axolotl brain and regeneration of the telencephalon.
Neural Dev. 2013 Jan 17;8:1.

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Rat anti BrdU antibody used for the identification of proliferating cells in the axolotl brain by immunohistochemistry on formalin fixed, paraffin embedded tissue sections.
Image caption:
Regeneration of the axolotl telencephalon. (A) Normal forebrain (ablated region marked with red box). (B) 15 weeks later showing complete regeneration. (C) RT97 wholemount showing the medial olfactory tract. (D) 9 weeks after removal of the right anterior third and olfactory bulb showing failure of regeneration and swelling of the regenerating olfactory nerve (arrowhead). (E) After removal of the olfactory bulb (right) the telencephalon heals but does not regenerate until the olfactory nerve regenerates (arrowhead = olfactory nerve swelling). (F-L) Regeneration after removal of the middle third of the telencephalon (as in A). After 3 weeks (F, G) there is no regeneration but an increase in BrdU+ cells in the damaged VZ (right). Close-up of the undamaged (H) and damaged (I) VZ shows more BrdU+ cells in I. J, BrdU+ counts in the VZ from 3-week regenerating (3r), 3-week undamaged (3c), 6-week regenerating (6r), 6-week undamaged (6c) with standard deviations. K, after 6 weeks there is restoration of the damaged side (right) but not full tissue replacement. After overnight BrdU labeling at 6-weeks there is local proliferation at the cut site (L). (M-Q) Regeneration after dorsal pallium removal (red box in M). M, damage site (right) after 8 days. N, after 24 days there is near complete regeneration. O, 40 days after removal the left and right telencephalons are indistinguishable (damage site marked with red star). P, regenerated brain 6 weeks after removal of the right dorsal pallium (as in M) shows complete regeneration. Q, 6 weeks after right dorsal pallium removal and olfactory nerve severing there is wound repair but shrinkage of the right telencephalon while the olfactory nerve regenerates. Red bars show the left/right difference in telencephalon length. Scale bars = 1 mm.

From: Maden M, Manwell LA, Ormerod BK. Proliferation zones in the axolotl brain and regeneration of the telencephalon.
Neural Dev. 2013 Jan 17;8:1.

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Rat anti BrdU antibody, clone Bu1/75 used for the detection of proliferating cells in the rat kidney by immunofluorescence on formalin fixed, paraffing embedded tissue sections.
Image caption:
Localization of BrdU-positive cells in the normal rat kidneys after long-term BrdU labeling. a–c BrdU was given intraperitoneally to normal rats for 4 weeks. Double staining of BrdU with several tubular (a), glomerular (b), and interstitial (c) markers was performed. Markers: aquaporin-1 (AQP-1), lotus tetragonolobus agglutinin (LTA), Tamm-Horsfall glycoprotein (THP), aquaporin-2 (AQP-2), alpha-SMA, vimentin, CD31, WT1, and aminopeptidase P. DAPI (blue). Magnification, ×1000. d Quantitative analysis of percentage of BrdU-positive cells per total marker-positive cells. Values are means?±?SE (n?=?5).

From: Nakasatomi M, Maeshima A, Mishima K, Ikeuchi H, Sakairi T, Kaneko Y, Hiromura K, Nojima Y. Novel approach for the detection of tubular cell migration into the interstitium during renal fibrosis in rats.
Fibrogenesis Tissue Repair. 2015 Jul 10;8:12.

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Rat anti BrdU antibody, clone Bu1/75 used for the detection of proliferating cells in the rat kidney by immunohistochemistry and immunofluorescence on formalin fixed, paraffing embedded tissue sections.
Image caption:
Migration of BrdU-positive tubular cells into the interstitium of the UUO kidneys. a Masson’s trichrome staining (a–d) and EVG staining (e–f) of kidney sections. (a, e) Normal kidney; (b, f) contralateral kidney, 10 days; (c, g) UUO kidney, 6 days; and (d, h) UUO kidney, 10 days. Magnification, ×200. b Localization of BrdU-positive cells in the normal (a), contralateral (b), and UUO (c–f) kidneys of rats with long-term BrdU labeling. Magnification, ×1000. Arrowheads indicate BrdU-positive cells protruding from TBM or migrating into interstitium. c Destruction of tubular basement membrane in the UUO kidneys of rats with long-term BrdU labeling. BrdU (red), markers (green), and DAPI (blue). Magnification, ×1000.

From: Nakasatomi M, Maeshima A, Mishima K, Ikeuchi H, Sakairi T, Kaneko Y, Hiromura K, Nojima Y. Novel approach for the detection of tubular cell migration into the interstitium during renal fibrosis in rats.
Fibrogenesis Tissue Repair. 2015 Jul 10;8:12.

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Rat anti BrdU antibody, clone Bu1/75 used for the detection of proliferating cells in the rat kidney by immunofluorescence on formalin fixed, paraffing embedded tissue sections.
Image caption:
Phenotypes of BrdU-positive tubular cells in the UUO kidneys. a Expression of myofibroblast-related markers in kidneys at 7 days after UUO. BrdU (red), markers (green), and DAPI (blue). Magnification, ×1000. Arrowheads indicate BrdU-positive cells coexpressing myofibroblast markers. b Number of alpha-SMA-positive cells and alpha-SMA/BrdU double-positive cells in the interstitium of contralateral and UUO kidneys at 8 days after operation. Values are means?±?SE (n?=?5). c Expression of AQP-1 in kidneys at 7 days after UUO. Arrowheads indicate BrdU-positive cells losing AQP-1 expression. BrdU (red), AQP-1 (green), and DAPI (blue). Magnification, ×1000. d Expression of inflammatory cell markers in kidneys at 7 days after UUO. DAPI (blue). Magnification, ×1000.

From: Nakasatomi M, Maeshima A, Mishima K, Ikeuchi H, Sakairi T, Kaneko Y, Hiromura K, Nojima Y. Novel approach for the detection of tubular cell migration into the interstitium during renal fibrosis in rats.
Fibrogenesis Tissue Repair. 2015 Jul 10;8:12.

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Rat anti BrdU antibody used for the detection of CldU by immunofluorescence demonstrating lack of reactivity with IdU
Image caption:
Cross reactivity of primary antibodies with the thymidine analogs was tested by incubation of sections of CldU only and IdU only treated animals with anti-IdU or anti-CldU antibodies, respectively (followed by incubation in the corresponding secondary antibodies.

From: Olivera-Pasilio V, Peterson DA, Castelló ME. Spatial distribution and cellular composition of adult brain proliferative zones in the teleost, Gymnotus omarorum.
Front Neuroanat. 2014 Sep 8;8:88.

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Rat anti BrdU antibody, clone Bu1/75 used for the detection of CldU in teleost brain by immunofluorescence
Image caption:
Cellular composition of adult G. omarorum brain proliferation zones revealed by the double thymidine analog labeling technique. (A,B) Projections of 25 confocal planes (sequentially acquired at one micron intervals) into a single plane of frontal sections at the level of the rombencephalic ventricle of fish treated with Protocol 1 (A) or 2 (B). (C,D) Orthogonal projections in the XZ and YZ axes, crossing at the position indicated by the yellow arrows in (A,B), respectively. As shown in (A,C), a high proportion of proliferating cells was double-labeled after a short chase (24 h) indicating cell cycle reentry, and thus correspond to fast cycling cells (yellow arrow). Single labeled cells correspond to actively cycling cells at the moment of IdU (green arrow) and CldU (red arrow) exposure. Double labeled cells in (B,D) reentered the cell cycle after a long chase (1 month) and thus correspond to slow cycling cells which were scarce (yellow arrow). Note the pleiotropic nuclear staining, with varying labeling intensity and surface aspect (either smooth or grainy). Scale bars: (A,B) = 50 μm; (C,D) = 5 μm. Generalized green staining at the brain surface in (B) is due to non-specific fluorescence of remaining embedding gelatin/albumin mixture.

From: Olivera-Pasilio V, Peterson DA, Castelló ME. Spatial distribution and cellular composition of adult brain proliferative zones in the teleost, Gymnotus omarorum.
Front Neuroanat. 2014 Sep 8;8:88.

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Rat anti BrdU antibody, clone Bu1/75 used for the detection of CldU in teleost brain by immunofluorescence
Image caption:
Telencephalic proliferation zones: location and cellular composition. Confocal images of frontal sections at rostral (A–B) intermediate (C) and caudal levels (D) of the telencephalic hemispheres and caudal levels of the olfactory bulb (A,B) of animals treated according to Protocol 1 (A) or 2 (B–D). Images in (A,A'–D,D') correspond to 30 sequentially acquired confocal planes (every 1 µm) projected to one plane. To corroborate the double labeling of nuclei, XZ and YZ orthogonal projections of the stacks were obtained at the sites indicated by the crossing lines (A?–D?). The location and boundaries of three telencephalic proliferation zones, determined by the spatial distribution of actively cycling cells IdU+ (green arrows in A',A?), CldU+ (red arrows in A'–D') and (A?–D?), were represented by arrowheads in (A–D) and dotted lines in (A'–D'): proliferation zone 1a at the free surface of the dorsal telencephalon; proliferation zone 1b at the ventral bulge of the telencephalic ventricle. Abundant double labeled nuclei were detected using Protocol 1 (fast cycling cells, yellow arrows in A',A?), while few double labeled nuclei were found using Protocol 2 (slow cycling cells, yellow arrows in B',D',B?,D?). Note that some cells that appeared to be double labeled in the projected images (white arrow in (C'), in fact corresponded to neighboring cells at the same x-y position but at different focal planes and separately labeled with only one of the thymidine analogs (C). Chains of elongated IdU+ or CldU+ nuclei are indicated by green and red double arrowheads, respectively. Long term weak analog label retaining cells are indicated by double green arrowheads in (C'–D'), indicating a long range radial migration. Scale bars: (A–D,A'–D') = 50 μm; (A?–D?) = 5 μm. DC, Central division of dorsal forebrain; DD, Dorsal division of the dorsal forebrain; DM, Dorsomedial telencephalon; DLd, Dorsolateral telencephalon, dorsal subdivision; V, Ventricle; Vd, Ventral telencephalon, dorsal subdivision; Vp, Ventral telencephalon, posterior subdivision; Vv, Ventral telencephalon, ventral subdivision.

From: Olivera-Pasilio V, Peterson DA, Castelló ME. Spatial distribution and cellular composition of adult brain proliferative zones in the teleost, Gymnotus omarorum.
Front Neuroanat. 2014 Sep 8;8:88.

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Rat anti BrdU antibody, clone Bu1/75 used for the detection of CldU in teleost brain by immunofluorescence
Image caption:
Mesencephalic proliferation zones: location and cellular composition of tectum opticum (TeO) and torus semicircularis (TS) proliferation zones. Confocal images of frontal sections at rostral (A,B) and caudal levels (C,D,G,H) levels of the mesencephalon of animals treated according to Protocol 1 (A,C,E,F) or 2 (B,D,G,H). Images in (A'–D',E–H) correspond to 30 sequentially acquired confocal planes (at 1 µm intervals) projected to one plane. To corroborate the double labeling of nuclei, XZ and YZ orthogonal projections of the stacks were obtained (A?,B?,E?–H?) at the sites indicated by the color coded arrows and the rectangles in the low power images. (A,B) The tectal commissure (cT) was flanked by two proliferation zones (arrowheads, A,B). Two groups of proliferating cells were evident in the dorsomedial and ventrolateral borders of the caudal part of the TeO (C',D',E,F) that converge into one at the caudal pole of the TeO (G). A single proliferation zone was evident at the ventromedial region of the TS (asterisks in C',D', F). Note the relative abundance of fast cycling cells in cT (A',A?) and TeO (E,E',F,F') and the scarcity of slow cycling cells in cT (B',B?) and the caudal pole of TeO (G',G?). A shift in the distribution of IdU + and CldU+ actively cycling cells was found in cT proliferation zone (B'). Scale bars: (A–D,G,H, A'–H') = 50 μm; (A?,B?,E?–H?) = 5 μm.

From: Olivera-Pasilio V, Peterson DA, Castelló ME. Spatial distribution and cellular composition of adult brain proliferative zones in the teleost, Gymnotus omarorum.
Front Neuroanat. 2014 Sep 8;8:88.

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Rat anti BrdU antibody, clone Bu1/75 used for the identificarion of dividing cells in mouse dentate gyrus by immunofluorescence.
Image caption:
Hippocampal cell proliferation, differentiation and survival in Tg mice with early AD-like pathology and age-matched Tg littermates.
(A) A representative image of the dentate gyrus stained for BrdU, demonstrates the distribution of proliferating BrdU+ cells. (B) A representative image of BrdU (green), DCX (red) and GFAP (white) positive cells in the dentate gyrus at 100 days of age. The arrow highlights a representative BrdU+/DCX+ cell. (C) A representative image of BrdU (green), NeuN (red) and GFAP (white) positive cells in the dentate gyrus. The arrow highlights a representative BrdU+/NeuN+ cell. Cell proliferation was examined at 100 days of age (D,E) while differentiation and survival was examined at 121 days of age (F,G) as a function of scyllo-inositol treatment. (D) The number of proliferating BrdU+ cells in the dentate gyrus of NTg (n = 7), NTg-SI (n = 6), Tg (n = 7) and Tg-SI (n = 5) mice were compared. This demonstrates more BrdU+ cells in Tg compared to NTg and to Tg-SI. (E) The percentage of BrdU+/DCX+ cells in the dentate gyrus showed less neuronal BrdU+ cells in Tg compared to the other three cohorts. (F) The number of BrdU+ cells surviving in NTg (n = 6), NTg-SI (n = 5), Tg (n = 6) and Tg-SI (n = 6) mice demonstrates more cell survival in Tg-SI mice. (G) The percentage of BrdU+/NeuN+ cells shows no difference between cohorts. Scale bars indicate 100 μm (A) or 25 μm (B,C). Data are mean ± SEM. One-way ANOVA with Fisher's Post-hoc test, * represents p<0.05 and ** represents p<0.01.

From: Morrone CD, Thomason LAM, Brown ME, Aubert I, McLaurin J (2016)
Effects of Neurotrophic Support and Amyloid-Targeted Combined Therapy on Adult Hippocampal Neurogenesis in a Transgenic Model of Alzheimer's Disease.
PLoS ONE 11(10): e0165393.

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Rat anti BrdU antibody, clone Bu1/75 used for the identificarion of dividing cells in mouse dentate gyrus by immunofluorescence.
Image caption:
Hippocampal cell proliferation in Tg mice with late AD-like pathology and age-matched NTg littermates.
Cell proliferation was assessed at 200 days of age (A,B). (A) The number of proliferating BrdU+ cells in NTg (n = 6), NTg-SI (n = 6), Tg (n = 6) and Tg-SI (n = 5) mice was compared, demonstrating less proliferation in Tg and Tg-SI vs. NTg-SI. (B) The percentage of BrdU+/DCX+ cells in the dentate gyrus was not different between cohorts. (C) Dentate gyrus stained with BrdU (red) and DCX (green) demonstrating the distribution of proliferating cells, as well as neuroblasts and immature neurons, respectively. (D) Representative orthogonal projection of a BrdU+/DCX+ cell. Scale bar indicates 100 μm (C) or 25 μm (D). Data are mean ± SEM. One-way ANOVA with Fisher's Post-hoc test, ** represents p<0.01.

From: Morrone CD, Thomason LAM, Brown ME, Aubert I, McLaurin J (2016)
Effects of Neurotrophic Support and Amyloid-Targeted Combined Therapy on Adult Hippocampal Neurogenesis in a Transgenic Model of Alzheimer's Disease.
PLoS ONE 11(10): e0165393.

Enlarge
BrdU Antibody | BU1/75 (ICR1) gallery image 26

Published customer image:
Rat anti BrdU antibody, clone Bu1/75 used for the identificarion of dividing cells in mouse dentate gyrus by immunofluorescence.
Image caption:
Hippocampal cell differentiation and survival in Tg mice with late AD-like pathology and age-matched NTg littermates.
Differentiation and survival was examined at 221 days of age (A,B) as a function of scyllo-inositol treatment. (A) The number of BrdU+ cells surviving in NTg (n = 7), NTg-SI (n = 7), Tg (n = 6) and Tg-SI (n = 6) mice showed no difference between treatment or genotype. (B) The percentage of BrdU+/NeuN+ cells in the dentate gyrus showed Tg and Tg-SI mice had a lower percentage of BrdU+/NeuN+ cells compared to NTg-SI mice. (C) Dentate gyrus stained with BrdU (red) and NeuN (blue) demonstrating the distribution of surviving newborn cells (red) within the population of mature granular neurons (blue). (D) Representative orthogonal projection of BrdU+/NeuN+ cell. Scale bar indicates 100 μm (C) or 25 μm (D). Data are mean ± SEM. One-way ANOVA with Fisher's Post-hoc test, * represents p<0.05 and ** represents p<0.01.

From: Morrone CD, Thomason LAM, Brown ME, Aubert I, McLaurin J (2016)
Effects of Neurotrophic Support and Amyloid-Targeted Combined Therapy on Adult Hippocampal Neurogenesis in a Transgenic Model of Alzheimer's Disease.
PLoS ONE 11(10): e0165393.

Enlarge
BrdU Antibody | BU1/75 (ICR1) gallery image 27

Proliferating cells stained with Rat anti BrdU antibody, clone Bu1/75 for newly synthesized DNA along with different intercalating (PI and 7-AAD) or minor groove binding (DAPI and Hoechst) dyes to stain total DNA, analysed by flow cytometry

Enlarge
BrdU Antibody | BU1/75 (ICR1) gallery image 28

Published customer image:
Rat anti BrdU antibody, clone Bu1/75 used for the identificarion of dividing cells in mouse dentate gyrus by immunofluorescence.
Image caption:
Hippocampal cell proliferation in scyllo-inositol, neotrofin, and scyllo-inositol/neotrofin treated 200 day old Tg mice.
(A) The number of proliferating BrdU+ cells in Tg-SI (n = 7), Tg-NEO (n = 6) and Tg-SI/NEO (n = 7) mice demonstrated no difference between cohorts. (B) The percentage of BrdU+/DCX+ cells (Tg-SI (n = 5), Tg-NEO (n = 6) and Tg-SI/NEO (n = 7)]) and (C) BrdU+/GFAP+ (Tg-SI (n = 7), Tg-NEO (n = 6) and Tg-SI/NEO (n = 7)) in the dentate gyrus of all groups were assessed and showed no changes between treatment groups. (D) Representative image of the dentate gyrus stained with BrdU (red) and GFAP (green) demonstrate the distribution of BrdU+ proliferating cells and astrocytes. (E) Representative BrdU+/GFAP+ cells are indicated by arrows. Scale bar indicates 100 μm (D) or 25 μm (E). Data are mean ± SEM. One-way ANOVA with Fisher's Post-hoc test.

From: Morrone CD, Thomason LAM, Brown ME, Aubert I, McLaurin J (2016)
Effects of Neurotrophic Support and Amyloid-Targeted Combined Therapy on Adult Hippocampal Neurogenesis in a Transgenic Model of Alzheimer's Disease.
PLoS ONE 11(10): e0165393.

Enlarge
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  • Rat anti BrdU
  • Rat anti BrdU:FITC
  • Rat anti BrdU:Biotin
  • Rat anti BrdU
  • Rat anti BrdU
  • Rat anti BrdU:FITC
  • Rat anti BrdU:Alexa Fluor® 488
(Rated 0.0 out of 5 based on 0 customer reviews)
    BrdU antibody, clone BU1/75 (ICR1) recognizes bromodeoxyuridine (known as BrdU or BrdUrd). The antibody reacts with BrdU incorporated into single stranded DNA, attached to a protein carrier and free BrdU. Bio-Rad offers 5 formats of the BrdU antibody.
    • Product Type
      Monoclonal Antibody
    • Clone
      BU1/75 (ICR1)
    • Isotype
      IgG2a
    4 Formats Available
      Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
      MCA2060BF *datasheet pdfdatasheet pdf0.1 mg
      MCA2060B
      MCA2060GAF *, IF, P *datasheet pdfdatasheet pdf0.1 mg
      MCA2060GA
      MCA2060FF *datasheet pdfdatasheet pdf0.1 mg
      MCA2060F
      MCA2060F *, IF, P *datasheet pdfdatasheet pdf0.2 mg
      MCA2060
      MCA2060FTF *datasheet pdfdatasheet pdf0.5 ml
      MCA2060FT
      MCA2060A488F *datasheet pdfdatasheet pdf100 Tests/1ml
      MCA2060A488
      MCA2060TF *, IF, P *datasheet pdfdatasheet pdf20 µg
      MCA2060T
      Summary
      Secondary Antibodies
      Negative Isotype Controls
      Useful Reagents
      Positive Controls
      Histology Controls
      More Images
      References
      Reviews
      -
      • Rat anti BrdU antibody clone BU1/75 (ICR1), recognizes bromodeoxyuridine (known as BrdU or BrdUrd). Rat anti BrdU antibody clone BU1/75 (ICR1) reacts with BrdU incorporated into single stranded DNA, attached to a protein carrier and free BrdU.

        Rat anti BrdU antibody, clone BU1/75 (ICR1) cross reacts with chlorodeoxyuridine (CldU) but does not cross react with thymidine or iododeoxyuridine (Aten et al. 1992). BrdU, IdU and CldU are analogs of thymidine, they can incorporate into DNA during DNA synthesis replacing thymidine.Antibody detection of incorporated BrdU in cellular DNA is extensively referenced as an accurate method to monitor cell proliferation in vivo and in vitro. In cell proliferation assays BrdU staining is coupled with the use of a dye that binds total DNA such as propidium iodide (PI). BrdU can be administered diluted in the culture medium or, in vivo via intraperitoneal injection, subcutaneous osmotic pump implants (Tesfaiqzi et al. 2004) or in drinking water (Moser et al. 2004)

        Clone BU1/75 (ICR1) has been used to detect CldU to study the speed of DNA replication fork (Bugler et al. 2010) , in the detection of CldU label retaining stem cells (Kimoto et al. 2008) and label retaining neurons (Murata et al. 2011).

      • Intended Use
      • Target Species
        Chemical
      • Product Form
        Purified IgG - liquid
        Purified IgG conjugated to Fluorescein Isothiocyanate Isomer 1 (FITC) - liquid
        Purified IgG conjugated to Biotin - liquid
        Purified IgG - liquid
        Purified IgG - liquid
        Purified IgG conjugated to Fluorescein Isothiocyanate Isomer 1 (FITC) - liquid
        Purified IgG conjugated to Alexa Fluor 488 - liquid
      • Reconstitution
      • Preparation
        Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
        Pack Size: 0.1 mgPurified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant.
        Pack Size: 0.5 mlPurified IgG prepared by affinity chromatography on Protein G
        Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
        Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
        Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
        Pack Size: 0.1 mgPurified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant.
        Pack Size: 0.5 mlPurified IgG prepared by affinity chromatography on Protein G
        Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
      • Preservative Stabilisers
        0.09% Sodium Azide (NaN3)
        Pack Size: 0.1 mg0.09% Sodium Azide (NaN3)
        1% Bovine Serum Albumin
        Pack Size: 0.5 ml0.09% Sodium Azide (NaN3)
        50% Glycerol
        0.09% Sodium Azide (NaN3)
        1% Bovine Serum Albumin
        0.09% Sodium Azide (NaN3)
        0.09% Sodium Azide (NaN3)
        Pack Size: 0.1 mg0.09% Sodium Azide (NaN3)
        1% Bovine Serum Albumin
        Pack Size: 0.5 ml0.09% Sodium Azide (NaN3)
        50% Glycerol
        0.09% Sodium Azide (NaN3)
        1% Bovine Serum Albumin
      • Purity
      • Approx. Protein Concentrations
        IgG concentration 1.0mg/ml
        Pack Size: 0.1 mgIgG concentration 0.1mg/ml
        Pack Size: 0.5 mlIgG concentration 0.5 mg/ml
        IgG concentration 0.1mg/ml
        IgG concentration 1.0mg/ml
        IgG concentration 1.0mg/ml
        Pack Size: 0.1 mgIgG concentration 0.1mg/ml
        Pack Size: 0.5 mlIgG concentration 0.5 mg/ml
        IgG concentration 0.05mg/ml
      • Reagents In The Kit
      • Preparing The Antibody
      • Test Principle
      • Buffer Solution
        Phosphate buffered saline
        Phosphate buffered saline
        Phosphate buffered saline
        Phosphate buffered saline
        Phosphate buffered saline
        Phosphate buffered saline
        Phosphate buffered saline
      • Storage
        Store at +4oC or at -20oC if preferred.

        This product should be stored undiluted.

        Storage in frost free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the protein. Should this product contain a precipitate we recommend microcentrifugation before use.
        Store at +4oC or at -20oC if preferred.

        This product should be stored undiluted.

        Storage in frost free freezers is not recommended. This product is photosensitive and should be protected from light.

        Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
        Store at +4oC or at -20oC if preferred.

        This product should be stored undiluted.

        Storage in frost free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the protein. Should this product contain a precipitate we recommend microcentrifugation before use.
        Store at +4oC or at -20oC if preferred.

        This product should be stored undiluted.

        Storage in frost free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the protein. Should this product contain a precipitate we recommend microcentrifugation before use.
        Store at +4oC or at -20oC if preferred.

        This product should be stored undiluted.

        Storage in frost free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the protein. Should this product contain a precipitate we recommend microcentrifugation before use.
        Store at +4oC or at -20oC if preferred.

        This product should be stored undiluted.

        Storage in frost free freezers is not recommended. This product is photosensitive and should be protected from light.

        Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
        Store at +4oC or at -20oC if preferred.
        Storage in frost-free freezers is not recommended.
        This product should be stored undiluted. This product is photosensitive and should be protected from light.
        Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
      • Shelf Life
        18 months from date of despatch.
        18 months from date of despatch.
        18 months from date of despatch.
        18 months from date of despatch.
        18 months from date of despatch.
        18 months from date of despatch.
        18 months from date of despatch.
      • Acknowledgements
        This product is provided under an intellectual property licence from Life Technologies Corporation. The transfer of this product is contingent on the buyer using the purchase product solely in research, excluding contract research or any fee for service research, and the buyer must not sell or otherwise transfer this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; (c) manufacturing or quality assurance or quality control, or (d) resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad CA 92008 USA or outlicensing@thermofisher.com
      • Regulatory
        For research purposes only
      • This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.

      • Application NameYesNoMin DilutionMax Dilution
        Immunofluorescence
        Flow Cytometry(1)1/251/200
        Immunohistology - Paraffin(2)1/251/200
        (1)
        See recommended protocol below.
        (2)
        See recommended protocol below.
      • Application NameYesNoMin DilutionMax Dilution
        Flow Cytometry(1)Neat
        1/5Pack Size: 0.1 mg
        1/20Pack Size: 0.5 ml
        (1)
        See recommended protocol below.
      • Application NameYesNoMin DilutionMax Dilution
        Flow Cytometry(1)Neat
        (1)
        See recommended protocol below.
      • Application NameYesNoMin DilutionMax Dilution
        Immunofluorescence
        Flow Cytometry(1)1/251/200
        Immunohistology - Paraffin(2)1/251/200
        (1)
        See recommended protocol below.
        (2)
        See recommended protocol below.
      • Application NameYesNoMin DilutionMax Dilution
        Immunofluorescence
        Flow Cytometry(1)1/251/200
        Immunohistology - Paraffin(2)1/251/200
        (1)
        See recommended protocol below.
        (2)
        See recommended protocol below.
      • Application NameYesNoMin DilutionMax Dilution
        Flow Cytometry(1)Neat
        1/5Pack Size: 0.1 mg
        1/20Pack Size: 0.5 ml
        (1)
        See recommended protocol below.
      • Application NameYesNoMin DilutionMax Dilution
        Flow Cytometry(1)Neat
        (1)
        See recommended protocol below.

      • Where this antibody has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the antibody for use in their own system using appropriate negative/positive controls.

      • Technical Advice
      • Technical Advice
      • Technical Advice
      • Technical Advice
      • Technical Advice
      • Technical Advice
      • Technical Advice
      • Recommended Protocol
        FLOW CYTOMETRY ANALYSIS

        Prepare the following solutions before proceeding:
        Phosphate buffered saline (PBS)
        2N HCl containing 0.5% Triton X-100
        PBS containing 0.05% Tween-20
        PBS containing 1% BSA (PBS/BSA)
        10mg/ml Propidium iodide (PI)
        0.1M Na2B4O7, pH 8.5

        1. Add BrdU to the cell suspension in culture medium to a final concentration of 10 µmol/L and incubate for 30 minutes in a CO2 incubator at 37°C.

        2. Wash cells twice with PBS/BSA by centrifuging at 500g for 10 minutes, decant supernatant and resuspend in a minimum volume of PBS.

        3. Add cells slowly into 5ml of 70% ethanol at -20°C, mixing continuously (vortex preferred).
        Incubate on ice for 30 minutes.

        4. Centrifuge at 500g for 10 minutes, decant supernatant, and resuspend cell pellet.

        5. Add 2ml of 2N HCl containing 0.5% Triton X-100 and incubate the cells for 30 minutes at room temperature (preferably on a rocking platform).

        6. Centrifuge at 500g for 10 minutes, decant supernatant and resuspend in 3 ml of 0.1M Na2B4O7, pH 8.5.

        7. Centrifuge at 500g for 10 minutes, decant supernatant and resuspend the cells in PBS/BSA + 0.05% Tween-20. Adjust cell concentration to 1 x 107/ml.

        8. Aliquot 100ul of cell suspension into required number of 12 x 75mm tubes.

        9. Incubate the cells with the BrdU antibody at the recommended dilution for 45 minutes at room temperature or overnight at 4°C.

        10. Add 2 ml of PBS/BSA and centrifuge the cells at 1000rpm for 5 minutes.

        11. If a secondary antibody layer is required then decant the supernatant and incubate the cells with the secondary antibody for 30 minutes at room temperature. If no secondary antibody layer is required then proceed to step 13.

        12. Wash the cells after the secondary antibody layer by repeating step 10.

        13. Decant the supernatant and add 1ml of PBS containing 10µg/ml PI (Dilute the 10mg/ml solution of PI 1/1000 in a suitable volume of PBS).

        14. Analyze cells by flow cytometry following the manufacturer’s instructions. The PI should be read on the appropriate channel set to the Peak/Area and not log scale.

        For Flow Cytometry references, please visit the following website:
        www.bio-rad-antibodies.com/support/brdu_antibody_clone_bu1_75_
        icr1_references-985.html


        IMMUNOHISTOLOGY

        Formalin-fixed paraffin-embedded tissue sections:

        Clone BU1/75 (ICR1) can be used for labeling paraffin-embedded tissue sections fixed in formalin. Denaturation of the DNA is critical for successful staining of BrdU. This can be achieved by exposing cells to heat, or acid. For heat-induced epitope retrieval, 10mM citrate buffer pH6.0 is recommended. Alternatively, a 30 min incubation in 2M HCl can be performed. The HCl must then be neutralized for 2 min with 0.1 M Na2B4O7. Pretreatment of tissues with proteinase K should be avoided.

        For Immunohistology references, please visit the following website:
        www.bio-rad-antibodies.com/support/brdu_antibody_clone_bu1_75_
        icr1_references-985.html
      • Recommended Protocol
        FLOW CYTOMETRY ANALYSIS

        Prepare the following solutions before proceeding:
        Phosphate buffered saline (PBS)
        2N HCl containing 0.5% Triton X-100
        PBS containing 0.05% Tween-20
        PBS containing 1% BSA (PBS/BSA)
        10mg/ml Propidium iodide (PI)
        0.1M Na2B4O7, pH 8.5

        1. Add BrdU to the cell suspension in culture medium to a final concentration of 10 µmol/L and incubate for 30 minutes in a CO2 incubator at 37°C.

        2. Wash cells twice with PBS/BSA by centrifuging at 500g for 10 minutes, decant supernatant and resuspend in a minimum volume of PBS.

        3. Add cells slowly into 5ml of 70% ethanol at -20°C, mixing continuously (vortex preferred).
        Incubate on ice for 30 minutes.

        4. Centrifuge at 500g for 10 minutes, decant supernatant, and resuspend cell pellet.

        5. Add 2ml of 2N HCl containing 0.5% Triton X-100 and incubate the cells for 30 minutes at room temperature (preferably on a rocking platform).

        6. Centrifuge at 500g for 10 minutes, decant supernatant and resuspend in 3 ml of 0.1M Na2B4O7, pH 8.5.

        7. Centrifuge at 500g for 10 minutes, decant supernatant and resuspend the cells in PBS/BSA + 0.05% Tween-20. Adjust cell concentration to 1 x 107/ml.

        8. Aliquot 100ul of cell suspension into required number of 12 x 75mm tubes.

        9. Incubate the cells with the BrdU antibody at the recommended dilution for 45 minutes at room temperature or overnight at 4°C.

        10. Add 2 ml of PBS/BSA and centrifuge the cells at 1000rpm for 5 minutes.

        11. If a secondary antibody layer is required then decant the supernatant and incubate the cells with the secondary antibody for 30 minutes at room temperature. If no secondary antibody layer is required then proceed to step 13.

        12. Wash the cells after the secondary antibody layer by repeating step 10.

        13. Decant the supernatant and add 1ml of PBS containing 10µg/ml PI (Dilute the 10mg/ml solution of PI 1/1000 in a suitable volume of PBS).

        14. Analyze cells by flow cytometry following the manufacturer’s instructions. The PI should be read on the appropriate channel set to the Peak/Area and not log scale.

        For Flow Cytometry references, please visit the following website:
        www.bio-rad-antibodies.com/support/brdu_antibody_clone_bu1_75_
        icr1_references-985.html
      • Recommended Protocol
        FLOW CYTOMETRY ANALYSIS

        Prepare the following solutions before proceeding:
        Phosphate buffered saline (PBS)
        2N HCl containing 0.5% Triton X-100
        PBS containing 0.05% Tween-20
        PBS containing 1% BSA (PBS/BSA)
        10mg/ml Propidium iodide (PI)
        0.1M Na2B4O7, pH 8.5

        1. Add BrdU to the cell suspension in culture medium to a final concentration of 10 µmol/L and incubate for 30 minutes in a CO2 incubator at 37°C.

        2. Wash cells twice with PBS/BSA by centrifuging at 500g for 10 minutes, decant supernatant and resuspend in a minimum volume of PBS.

        3. Add cells slowly into 5ml of 70% ethanol at -20°C, mixing continuously (vortex preferred).
        Incubate on ice for 30 minutes.

        4. Centrifuge at 500g for 10 minutes, decant supernatant, and resuspend cell pellet.

        5. Add 2ml of 2N HCl containing 0.5% Triton X-100 and incubate the cells for 30 minutes at room temperature (preferably on a rocking platform).

        6. Centrifuge at 500g for 10 minutes, decant supernatant and resuspend in 3 ml of 0.1M Na2B4O7, pH 8.5.

        7. Centrifuge at 500g for 10 minutes, decant supernatant and resuspend the cells in PBS/BSA + 0.05% Tween-20. Adjust cell concentration to 1 x 107/ml.

        8. Aliquot 100ul of cell suspension into required number of 12 x 75mm tubes.

        9. Incubate the cells with the BrdU antibody at the recommended dilution for 45 minutes at room temperature or overnight at 4°C.

        10. Add 2 ml of PBS/BSA and centrifuge the cells at 1000rpm for 5 minutes.

        11. If a secondary antibody layer is required then decant the supernatant and incubate the cells with the secondary antibody for 30 minutes at room temperature. If no secondary antibody layer is required then proceed to step 13.

        12. Wash the cells after the secondary antibody layer by repeating step 10.

        13. Decant the supernatant and add 1ml of PBS containing 10µg/ml PI (Dilute the 10mg/ml solution of PI 1/1000 in a suitable volume of PBS).

        14. Analyze cells by flow cytometry following the manufacturer’s instructions. The PI should be read on the appropriate channel set to the Peak/Area and not log scale.

        For Flow Cytometry references, please visit the following website:
        www.bio-rad-antibodies.com/support/brdu_antibody_clone_bu1_75_
        icr1_references-985.html
      • Recommended Protocol
        FLOW CYTOMETRY ANALYSIS

        Prepare the following solutions before proceeding:
        Phosphate buffered saline (PBS)
        2N HCl containing 0.5% Triton X-100
        PBS containing 0.05% Tween-20
        PBS containing 1% BSA (PBS/BSA)
        10mg/ml Propidium iodide (PI)
        0.1M Na2B4O7, pH 8.5

        1. Add BrdU to the cell suspension in culture medium to a final concentration of 10 µmol/L and incubate for 30 minutes in a CO2 incubator at 37°C.

        2. Wash cells twice with PBS/BSA by centrifuging at 500g for 10 minutes, decant supernatant and resuspend in a minimum volume of PBS.

        3. Add cells slowly into 5ml of 70% ethanol at -20°C, mixing continuously (vortex preferred).
        Incubate on ice for 30 minutes.

        4. Centrifuge at 500g for 10 minutes, decant supernatant, and resuspend cell pellet.

        5. Add 2ml of 2N HCl containing 0.5% Triton X-100 and incubate the cells for 30 minutes at room temperature (preferably on a rocking platform).

        6. Centrifuge at 500g for 10 minutes, decant supernatant and resuspend in 3 ml of 0.1M Na2B4O7, pH 8.5.

        7. Centrifuge at 500g for 10 minutes, decant supernatant and resuspend the cells in PBS/BSA + 0.05% Tween-20. Adjust cell concentration to 1 x 107/ml.

        8. Aliquot 100ul of cell suspension into required number of 12 x 75mm tubes.

        9. Incubate the cells with the BrdU antibody at the recommended dilution for 30 minutes at room temperature.

        10. Add 2 ml of PBS/BSA and centrifuge the cells at 1000rpm for 5 minutes.

        11. If a secondary antibody layer is required then decant the supernatant and incubate the cells with the secondary antibody for 30 minutes at room temperature. If no secondary antibody layer is required then proceed to step 13.

        12. Wash the cells after the secondary antibody layer by repeating step 10.

        13. Decant the supernatant and add 1ml of PBS containing 10µg/ml PI (Dilute the 10mg/ml solution of PI 1/1000 in a suitable volume of PBS).

        14. Analyze cells by flow cytometry following the manufacturer’s instructions. The PI should be read on the appropriate channel set to the Peak/Area and not log scale.

        For Flow Cytometry references, please visit the following website:
        www.bio-rad-antibodies.com/support/brdu_antibody_clone_bu1_75_
        icr1_references-985.html


        IMMUNOHISTOLOGY

        Formalin-fixed paraffin-embedded tissue sections:

        Clone BU1/75 (ICR1) can be used for labeling paraffin-embedded tissue sections fixed in formalin. Denaturation of the DNA is critical for successful staining of BrdU. This can be achieved by exposing cells to heat, or acid. For heat-induced epitope retrieval, 10mM citrate buffer pH6.0 is recommended. Alternatively, a 30 min incubation in 2M HCl can be performed. The HCl must then be neutralized for 2 min with 0.1 M Na2B4O7. Pretreatment of tissues with proteinase K should be avoided.

        For Immunohistology references, please visit the following website:
        www.bio-rad-antibodies.com/support/brdu_antibody_clone_bu1_75_
        icr1_references-985.html
      • Recommended Protocol
        FLOW CYTOMETRY ANALYSIS

        Prepare the following solutions before proceeding:
        Phosphate buffered saline (PBS)
        2N HCl containing 0.5% Triton X-100
        PBS containing 0.05% Tween-20
        PBS containing 1% BSA (PBS/BSA)
        10mg/ml Propidium iodide (PI)
        0.1M Na2B4O7, pH 8.5

        1. Add BrdU to the cell suspension in culture medium to a final concentration of 10 µmol/L and incubate for 30 minutes in a CO2 incubator at 37°C.

        2. Wash cells twice with PBS/BSA by centrifuging at 500g for 10 minutes, decant supernatant and resuspend in a minimum volume of PBS.

        3. Add cells slowly into 5ml of 70% ethanol at -20°C, mixing continuously (vortex preferred).
        Incubate on ice for 30 minutes.

        4. Centrifuge at 500g for 10 minutes, decant supernatant, and resuspend cell pellet.

        5. Add 2ml of 2N HCl containing 0.5% Triton X-100 and incubate the cells for 30 minutes at room temperature (preferably on a rocking platform).

        6. Centrifuge at 500g for 10 minutes, decant supernatant and resuspend in 3 ml of 0.1M Na2B4O7, pH 8.5.

        7. Centrifuge at 500g for 10 minutes, decant supernatant and resuspend the cells in PBS/BSA + 0.05% Tween-20. Adjust cell concentration to 1 x 107/ml.

        8. Aliquot 100ul of cell suspension into required number of 12 x 75mm tubes.

        9. Incubate the cells with the BrdU antibody at the recommended dilution for 45 minutes at room temperature or overnight at 4°C.

        10. Add 2 ml of PBS/BSA and centrifuge the cells at 1000rpm for 5 minutes.

        11. If a secondary antibody layer is required then decant the supernatant and incubate the cells with the secondary antibody for 30 minutes at room temperature. If no secondary antibody layer is required then proceed to step 13.

        12. Wash the cells after the secondary antibody layer by repeating step 10.

        13. Decant the supernatant and add 1ml of PBS containing 10µg/ml PI (Dilute the 10mg/ml solution of PI 1/1000 in a suitable volume of PBS).

        14. Analyze cells by flow cytometry following the manufacturer’s instructions. The PI should be read on the appropriate channel set to the Peak/Area and not log scale.

        For Flow Cytometry references, please visit the following website:
        www.bio-rad-antibodies.com/support/brdu_antibody_clone_bu1_75_
        icr1_references-985.html


        IMMUNOHISTOLOGY

        Formalin-fixed paraffin-embedded tissue sections:

        Clone BU1/75 (ICR1) can be used for labeling paraffin-embedded tissue sections fixed in formalin. Denaturation of the DNA is critical for successful staining of BrdU. This can be achieved by exposing cells to heat, or acid. For heat-induced epitope retrieval, 10mM citrate buffer pH6.0 is recommended. Alternatively, a 30 min incubation in 2M HCl can be performed. The HCl must then be neutralized for 2 min with 0.1 M Na2B4O7. Pretreatment of tissues with proteinase K should be avoided.

        For Immunohistology references, please visit the following website:
        www.bio-rad-antibodies.com/support/brdu_antibody_clone_bu1_75_
        icr1_references-985.html
      • Recommended Protocol
        FLOW CYTOMETRY ANALYSIS

        Prepare the following solutions before proceeding:
        Phosphate buffered saline (PBS)
        2N HCl containing 0.5% Triton X-100
        PBS containing 0.05% Tween-20
        PBS containing 1% BSA (PBS/BSA)
        10mg/ml Propidium iodide (PI)
        0.1M Na2B4O7, pH 8.5

        1. Add BrdU to the cell suspension in culture medium to a final concentration of 10 µmol/L and incubate for 30 minutes in a CO2 incubator at 37°C.

        2. Wash cells twice with PBS/BSA by centrifuging at 500g for 10 minutes, decant supernatant and resuspend in a minimum volume of PBS.

        3. Add cells slowly into 5ml of 70% ethanol at -20°C, mixing continuously (vortex preferred).
        Incubate on ice for 30 minutes.

        4. Centrifuge at 500g for 10 minutes, decant supernatant, and resuspend cell pellet.

        5. Add 2ml of 2N HCl containing 0.5% Triton X-100 and incubate the cells for 30 minutes at room temperature (preferably on a rocking platform).

        6. Centrifuge at 500g for 10 minutes, decant supernatant and resuspend in 3 ml of 0.1M Na2B4O7, pH 8.5.

        7. Centrifuge at 500g for 10 minutes, decant supernatant and resuspend the cells in PBS/BSA + 0.05% Tween-20. Adjust cell concentration to 1 x 107/ml.

        8. Aliquot 100ul of cell suspension into required number of 12 x 75mm tubes.

        9. Incubate the cells with the BrdU antibody at the recommended dilution for 30 minutes at room temperature.

        10. Add 2 ml of PBS/BSA and centrifuge the cells at 1000rpm for 5 minutes.

        11. If a secondary antibody layer is required then decant the supernatant and incubate the cells with the secondary antibody for 30 minutes at room temperature. If no secondary antibody layer is required then proceed to step 13.

        12. Wash the cells after the secondary antibody layer by repeating step 10.

        13. Decant the supernatant and add 1ml of PBS containing 10µg/ml PI (Dilute the 10mg/ml solution of PI 1/1000 in a suitable volume of PBS).

        14. Analyze cells by flow cytometry following the manufacturer’s instructions. The PI should be read on the appropriate channel set to the Peak/Area and not log scale.

        For Flow Cytometry references, please visit the following website:
        www.bio-rad-antibodies.com/support/brdu_antibody_clone_bu1_75_
        icr1_references-985.html
      • Recommended Protocol
        FLOW CYTOMETRY ANALYSIS

        Prepare the following solutions before proceeding:
        Phosphate buffered saline (PBS)
        2N HCl containing 0.5% Triton X-100
        PBS containing 0.05% Tween-20
        PBS containing 1% BSA (PBS/BSA)
        10mg/ml Propidium iodide (PI)
        0.1M Na2B4O7, pH 8.5

        1. Add BrdU to the cell suspension in culture medium to a final concentration of 10 µmol/L and incubate for 30 minutes in a CO2 incubator at 37°C.

        2. Wash cells twice with PBS/BSA by centrifuging at 500g for 10 minutes, decant supernatant and resuspend in a minimum volume of PBS.

        3. Add cells slowly into 5ml of 70% ethanol at -20°C, mixing continuously (vortex preferred).
        Incubate on ice for 30 minutes.

        4. Centrifuge at 500g for 10 minutes, decant supernatant, and resuspend cell pellet.

        5. Add 2ml of 2N HCl containing 0.5% Triton X-100 and incubate the cells for 30 minutes at room temperature (preferably on a rocking platform).

        6. Centrifuge at 500g for 10 minutes, decant supernatant and resuspend in 3 ml of 0.1M Na2B4O7, pH 8.5.

        7. Centrifuge at 500g for 10 minutes, decant supernatant and resuspend the cells in PBS/BSA + 0.05% Tween-20. Adjust cell concentration to 1 x 107/ml.

        8. Aliquot 100ul of cell suspension into required number of 12 x 75mm tubes.

        9. Incubate the cells with the BrdU antibody at the recommended dilution for 30 minutes at room temperature.

        10. Add 2 ml of PBS/BSA and centrifuge the cells at 1000rpm for 5 minutes.

        11. If a secondary antibody layer is required then decant the supernatant and incubate the cells with the secondary antibody for 30 minutes at room temperature. If no secondary antibody layer is required then proceed to step 13.

        12. Wash the cells after the secondary antibody layer by repeating step 10.

        13. Decant the supernatant and add 1ml of PBS containing 10µg/ml PI (Dilute the 10mg/ml solution of PI 1/1000 in a suitable volume of PBS).

        14. Analyze cells by flow cytometry following the manufacturer’s instructions. The PI should be read on the appropriate channel set to the Peak/Area and not log scale.

        For Flow Cytometry references, please visit the following website:
        www.bio-rad-antibodies.com/support/brdu_antibody_clone_bu1_75_
        icr1_references-985.html
      • Flow Cytometry
        Use 20ul of the suggested working dilution to label 106 cells in 100ul.
      • ELISA
      • ELISA
      • ELISA
      • ELISA
      • ELISA
      • ELISA
      • ELISA
      • Immunohistology
      • Immunohistology
      • Immunohistology
      • Immunohistology
      • Immunohistology
      • Immunohistology
      • Immunohistology
      • Histology Positive Control Tissue
      • Histology Positive Control Tissue
      • Histology Positive Control Tissue
      • Histology Positive Control Tissue
      • Histology Positive Control Tissue
      • Histology Positive Control Tissue
      • Histology Positive Control Tissue
      • Immunofluorescence
      • Immunofluorescence
      • Immunofluorescence
      • Immunofluorescence
      • Immunofluorescence
      • Immunofluorescence
      • Immunofluorescence
      • Western Blotting
      • Western Blotting
      • Western Blotting
      • Western Blotting
      • Western Blotting
      • Western Blotting
      • Western Blotting
      • Instructions For Use
      • Instructions For Use
      • Instructions For Use
      • Instructions For Use
      • Instructions For Use
      • Instructions For Use
      • Instructions For Use

      Additional BrdU Antibody Formats

      Formats Clone Applications Sizes available
      BrdU Antibody : FITC BU1/75 (ICR1) F * 0.5 ml | 0.1 mg
      BrdU Antibody : Purified BU1/75 (ICR1) F *, IF, P * 20 µg | 0.1 mg | 0.2 mg
      BrdU Antibody : Biotin BU1/75 (ICR1) F * 0.1 mg
      BrdU Antibody : Alexa Fluor® 488 BU1/75 (ICR1) F * 100 Tests/1ml
      • Copyright © 2016 Bio-Rad Antibodies (formerly AbD Serotec)

      Recommended Secondary Antibody

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        Goat F(ab')2 anti Rat IgG:FITC (Mouse Adsorbed)STAR690.5 mlF
        STAR69
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        STAR20A
        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
        Streptavidin:APCSTAR119100 TestsF
        STAR119
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        Goat anti Rat IgG:Alk. Phos. (Mouse Adsorbed)STAR131A1 mlC, E, P, WB
        STAR131A
        Goat anti Rat IgG:Biotin (Mouse Adsorbed)STAR131B0.5 mgC, E, IF, P, WB
        STAR131B
        Goat anti Rat IgG:DyLight®549 (Mouse Adsorbed)STAR71D549GA0.1 mgF, IF
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        Rabbit F(ab')2 anti Rat IgG:Dylight®800STAR16D800GA0.1 mgF, IF, WB
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        STAR71D800GA
        Goat F(ab')2 anti Rat IgG:FITC (Mouse Adsorbed)STAR690.5 mlF
        STAR69
        Rabbit F(ab')2 anti Rat IgG:FITCSTAR17B1 mgF
        STAR17B
        Goat anti Rat IgG:HRP (Mouse Adsorbed)STAR720.5 mgC, E, P
        STAR72
        Rabbit F(ab')2 anti Rat IgG:HRPSTAR21B1 mgC, E, P, RE
        STAR21B
        Goat F(ab')2 anti Rat IgG:RPE (Mouse Adsorbed)STAR730.5 mlF
        STAR73
        Rabbit F(ab')2 anti Rat IgG:RPESTAR20A1 mlF
        STAR20A
        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
        Rabbit F(ab')2 anti Rat IgG:FITCSTAR17B1 mgF
        STAR17B
        Rabbit F(ab')2 anti Rat IgG:HRPSTAR21B1 mgC, E, P, RE
        STAR21B
        Rabbit F(ab')2 anti Rat IgG:RPESTAR20A1 mlF
        STAR20A

        Recommended Negative Isotype Control

          DescriptionProduct CodePack SizeApplicationsList PriceQuantity
          Rat IgG2a Negative Control:Alexa Fluor® 488MCA1124A488100 Tests/1mlF
          MCA1124A488

          Useful Reagents

            Recommended Positive Controls

              Histology Controls

                References

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                BrdU Staining Protocol

                Recommended guidance for staining with Mouse anti-BrdU antibody for flow cytometry and immunohistochemistry on paraffin-embedded tissue sections fixed in formalin.

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