BrdU Antibody | BU1/75 (ICR1)

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Formalin fixed, paraffin embedded mouse colon stained with Rat anti BrdU (OBT0030): low power

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

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

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Formalin fixed, paraffin embedded mouse colon stained with Rat anti BrdU: high 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 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 used for the detection of BrdU positive cells in Mouse brain by immunohistochemistry on cryosections.
Image caption:
PEDF Treatment Does Not Affect Cell Proliferation After Striatal Ischemic Injury. Mice received daily injections of 50 mg/kg BrdU for seven days starting at the day of MCAO. (A) Representative images of cortex, striatum, subventricular zone (SVZ), and dentate gyrus of CSF and PEDF group. The dotted line in the images of SVZ indicates the area used for counting the cells (distance of 50 µm from the ventricles). Scale bar = 200 µm. (B–C) Graphs represent the number of BrdU-positive cells/mm2 in striatum, cortex, SVZ, and dentate gyrus. We did not find any significant differences between CSF and PEDF groups. (B) We found a significant difference between ipsilateral and contralateral striatum for both PEDF (repeated measures ANOVA, F(1,7) = 76.994, p<0.001) and CSF (F(1,7) = 96.351, p<0.001). However, for cortex, there is a significant difference between ipsilateral and contralateral cortex for PEDF (repeated measures ANOVA, F(1,7) = 14.415, p = 0.007), but not for CSF (F(1,7) = 3.016, p = .124). (C) We did not find any significant differences between ipsilateral and contralateral SVZ and dentate gyrus. Data is represented as means ± SD. n = 5 for CSF, n = 4 for PEDF.

From: Zille M, Riabinska A, Terzi MY, Balkaya M, Prinz V, et al. (2014) Influence of Pigment Epithelium-Derived Factor on Outcome after Striatal Cerebral Ischemia in the Mouse. PLoS ONE 9(12): e114595.

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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 positive cells in mouse brain by immunofluorescence.
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 used for the detection of BrdU positive cells in mouse retina by immunofluorescence.
Image caption:
Retinal progenitor cell (RPC) cell cycle is lengthened in the Ccnd1-/- retina. P0 wild-type and Ccnd1-/- retinas, cultured successively in iododeoxyuridine (IdU) for 2 hours and bromodeoxyuridine (BrdU) for 30 minutes, were triple-stained with mouse (A, D) anti-BrdU antibody (Ab) recognizing both IdU and BrdU, (B, E) rat anti-BrdU antibody recognizing only BrdU, and (C, F) with an antibody against PCNA marking RPCs. Arrows in (A-C) mark IdU+ only RPCs (IdU+, BrdU-; positive signal in (A, C) but not (B)) in wild-type retina that have moved up to the apical surface during the labeling period. Arrows in (D-F) mark IdU+ only RPCs (positive signal in (D, F) but not (E)) in the Ccnd1-/- retina during the same period. (G) Quantification of average RPC cell cycle time (Tc), S phase time (Ts) and G1 + G2 + M phase time (Tc - Ts) in wild-type (wt) and Ccnd1-/- retinas at E14.5 and P0. Scale bar: 50 µm; (F) is representative for (A-F).

From: Das G, Choi Y, Sicinski P, Levine EM. Cyclin D1 fine-tunes the neurogenic output of embryonic retinal progenitor cells.
Neural Dev. 2009 May 5;4:15.

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Rat anti BrdU antibody, clone Bu1/75 used for the detection of dividing cells in 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):1.

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Rat anti BrdU antibody, clone Bu1/75 used for the detection of dividing cells in 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):1.

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Rat anti BrdU antibody, clone Bu1/75 used for the detection of dividing cells in 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):1.

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Rat anti BrdU antibody, clone Bu1/75 used for the detection of dividing cells in 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):1.

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Published customer image:
Rat anti BrdU antibody, clone Bu1/75 used for the detection of dividing cells in axolotl brain by immunofluorescence 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):1.

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Published customer image:
Rat anti BrdU antibody, clone Bu1/75 used for the detection of dividing cells in 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):1.

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Rat anti BrdU antibody, clone Bu1/75 used for the identification of newly divided cells in rat jejenum 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. Intestinal epithelial stem/progenitor cells are controlled by mucosal afferent nerves.
PLoS One. 2011 Feb 9;6(2):e16295.

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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

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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.

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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.

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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.

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  • Rat anti BrdU
  • Rat anti BrdU:FITC
  • Rat anti BrdU
  • Rat anti BrdU
  • Rat anti BrdU
(Rated 0.0 out of 5 based on 0 customer reviews)
  • Product Type
    Monoclonal Antibody
  • Clone
    BU1/75 (ICR1)
  • Isotype
    IgG2a
4 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    OBT0030GF *, IF, P *datasheet pdfdatasheet pdf0.25 mg
    OBT0030G
    OBT0030CXF *, IF, P *datasheet pdfdatasheet pdf0.25 ml
    OBT0030CX
    OBT0030F *, IF, P *datasheet pdfdatasheet pdf0.5 mg
    OBT0030
    OBT0030FF *datasheet pdfdatasheet pdf1 ml
    OBT0030F
    OBT0030SF *, IF, P *datasheet pdfdatasheet pdf5 ml
    OBT0030S
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
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    • 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 FITC- liquid
      Purified IgG - liquid
      Tissue Culture Supernatant - liquid
      Ascites
    • Reconstitution
    • Preparation
      Purified IgG prepared by affinity chromatography on Protein G
      Purified IgG prepared by affinity chromatography on Protein G
      Purified IgG prepared by affinity chromatography on Protein G
      Tissue Culture Supernatant containing 5-10% foetal calf serum
    • Preservative Stabilisers
      0.09%Sodium Azide
      25%Glycerol
      0.09%Sodium Azide
      50%Glycerol
      0.09%Sodium Azide
      25%Glycerol
      0.09% Sodium Azide (NaN3)
      0.09%Sodium Azide
    • Purity
    • Approx. Protein Concentrations
      IgG concentration 0.5 mg/ml
      IgG concentration 0.5 mg/ml
      IgG concentration 0.5 mg/ml
    • Reagents In The Kit
    • Preparing The Antibody
    • Test Principle
    • Buffer Solution
      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.
      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.
      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.
    • 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.
    • Acknowledgements
    • 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)Neat1/20
      (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)neat1/20
      Immunohistology - Paraffin(2)neat1/10
      (1)
      See recommended protocol below.
      (2)
      See recommended protocol below.
    • Application NameYesNoMin DilutionMax Dilution
      Immunofluorescence
      Flow Cytometry(1)1/5001/2000
      Immunohistology - Paraffin(2)1/501/2000
      (1)
      See recommended protocol below.
      (2)
      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
    • 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 miniumum 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 by repeating step 10.

      13. Decant off 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. Analyse 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, pH8.5

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

      11. 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).

      12. 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 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 minimal 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 3ml 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 2ml 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 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, pH8.5

      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 3ml 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 2ml 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 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
    • Flow Cytometry
      Use 20ul of the suggested working dilution to label 106 cells in 100ul.
    • ELISA
    • ELISA
    • ELISA
    • ELISA
    • ELISA
    • 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
    • Immunofluorescence
    • Immunofluorescence
    • Immunofluorescence
    • Immunofluorescence
    • Immunofluorescence
    • 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

    Additional BrdU Antibody Formats

    Formats Clone Applications Sizes available
    BrdU Antibody : S/N BU1/75 (ICR1) F *, IF, P * 5 ml
    BrdU Antibody : Purified BU1/75 (ICR1) F *, IF, P * 0.5 mg | 0.25 mg
    BrdU Antibody : FITC BU1/75 (ICR1) F * 1 ml
    BrdU Antibody : Ascites BU1/75 (ICR1) F *, IF, P * 0.25 ml
    • Copyright © 2016 Bio-Rad Antibodies (formerly AbD Serotec)

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      Recommended Negative Isotype Control

        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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