CD45 Antibody | IBL-3/16

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CD45 Antibody | IBL-3/16 gallery image 1

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Rat anti Mouse CD45 antibody, clone IBL-3/16 used for the detection of mouse leukocytes by flow cytometry.
Image caption:
Infiltration of GFP+ BM-cells in infarct and peri-infarct regions. (A-B) Dot plots of viable macrophages/granulocytes (CD11b+CD45high, top right quadrants) and microglia (CD11b+CD45dim, bottom right quadrants) in cortex from BM-chimeric unmanipulated mice and mice exposed to pMCAO. (C) Bar graph showing mean numbers of CD11b+CD45dim microglia and CD11b+CD45high macrophages/granulocytes in BM-chimeric mice 24 hours after pMCAO, subdivided based on expression of GFP (n = 5). Approximately 92% of of the CD45high population were GFP+ . (D) Estimation and comparison of mean numbers of CD11b+CD45dim microglia in non-chimeric (n = 10) versus BM-chimeric mice (n = 5) 24 hours after of pMCAO shows significantly fewer CD11b+CD45dim microglial cells in irradiated mice. (E) Overview, showing distribution of infiltrating GFP+ BM-derived cells into infarct (IF) and peri-infarct (P-IF) regions 24 hours after pMCAO. (E-G) By 24 hours, GFP+ single cells (F) and vessel-associated aggregates of GFP+ cells (arrows in G) were observed in infarct and peri-infarct regions. Some of the vessel-associated cells were round, leukocyte-like cells (arrows) while others were elongated cells lining the vasculature (arrow heads in G and in insert). (H) Bar graph showing mean numbers of single GFP+ cells and vessel-associated aggregates of GFP+ cells in ipsi- and contralateral cortex 24 hours after surgery (n = 10). (I-P) Immunohistochemical staining of CD45.1 (I, K), CD45.2 (J, L), IgG2a (M, O) and CD45 (N, P) in ischemic tissue in BM-chimeric (I, J, M, N) and non-chimeric mice (K, L, O, P) 24 hours after pMCAO. N.D, none detected. Scale bars: 200 µm (A), 10 µm (B, C). 50 µm (I-P) *P < 0.05, **P < 0.01, and ***P < 0.001.

From: Clausen BH, Lambertsen KL, Babcock AA, Holm TH, Dagnaes-Hansen F, Finsen B. Interleukin-1beta and tumor necrosis factor-alpha are expressed by different subsets of microglia and macrophages after ischemic stroke in mice.
J Neuroinflammation. 2008 Oct 23;5:46.

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Rat anti Mouse CD45 antibody, clone IBL-3/16 used for the detection of murine leukocytes by flow cytometry.
Image caption:
Inflammatory response following permanent MCA occlusion. (A-C) Dot plots of viable CD11b+CD45high macrophages/granulocytes (top right quadrants) and CD11b+CD45dim microglia (bottom right quadrants) in cortex from unmanipulated control mice (A, B), and mice exposed to pMCAO with 24 hour survival (C). (D) At 24 hours, flow cytometric analysis of the CD11b+CD45high profiles showed that approximately half of the population consisted of CD45highGr1+ granulocytes. (E) Quantification of CD11b+CD45dim and CD11b+CD45high cells in unmanipulated control mice (n = 10), in mice 6 (n = 7), 12 (n = 7), or 24 hours after pMCAO (n = 10), and in sham-operated mice 24 hours after pMCAO (n = 7). (F) Bar graphs showing equal recruitment of CD11b+CD45highGr1- macrophages and CD11b-CD45highGr1+ granulocytes in unmanipulated mice, in mice 6, 12, or 24 hours after pMCAO, and in sham-operated mice 24 hours after pMCAO. (G, H) Bar graphs showing the mean fluorescent intensity (MFI) of CD45 expression by CD45dim microglia (G) and CD45high macrophages/granulocytes (H). *P < 0.05, **P < 0.01, and ***P < 0.001.
J Neuroinflammation. 2008 Oct 23;5:46. doi: 10.1186/1742-2094-5-46.

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Published customer image:
Rat anti Mouse CD45 antibody, clone IBL-3/16 used for the detection of murine leukocytes by flow cytometry.
Image caption:
Cytokine expression in segregated populations of cells following stroke. (A, B) Dot plots showing CD11b+CD45high macrophages/granulocytes (upper right quadrants) and CD11b+CD45dim microglia (bottom right quadrants) expressing IL-1β (A) or TNF-α (B). (C-J) Bar graphs showing numbers and proportions of IL-1β (C, D), TNF-α (F, G) and IL-1β/TNF-α co-expressing (I, J) CD11b+CD45dim microglia and CD11b+CD45high macrophages/granulocytes in unmanipulated control mice (n = 10), in mice 6 (n = 7), 12 (n = 7), or 24 hours after pMCAO (n = 10), and in sham-operated mice 24 hours after pMCAO (n = 7). (E, H) Comparison of the MFI values for IL-1β (E) and TNF-α (H) in viable CD11b+CD45dim microglia and CD11b+CD45high macrophages/granulocytes in unmanipulated mice, in mice 6, 12, or 24 hours after pMCAO, and in sham-operated mice 24 hours after pMCAO. Macrophages/granulocytes express significantly more IL-1β than do microglial in unmanipulated mice, in mice 6, 12, or 24 hours after pMCAO, and in sham-operated mice 24 hours after pMCAO (E), whereas microglial cells express significantly higher levels of TNF-α than do macrophages/granulocytes at 12 h and 24 hours, and in sham-operated mice 24 hours after pMCAO (H). (K) CD11b+CD45highGr1- macrophages and not CD11b+CD45highGr1+ granulocytes are the main producers of IL-1β and TNF-α 24 hours after pMCAO. *P < 0.05, **P < 0.01, and ***P < 0.001.

From: Clausen BH, Lambertsen KL, Babcock AA, Holm TH, Dagnaes-Hansen F, Finsen B. Interleukin-1beta and tumor necrosis factor-alpha are expressed by different subsets of microglia and macrophages after ischemic stroke in mice.
J Neuroinflammation. 2008 Oct 23;5:46. doi: 10.1186/1742-2094-5-46.

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Published customer image:
Rat anti Mouse CD45 antibody, clone IBL-3/16 used for the detection of leukocytes by flow cytometry.
Image caption:
Sensitivity of cytokine detection using flow cytometry. Histograms and dot plots of IL-1β (A) and TNF-α (B) expression in LPS-activated peritoneal macrophages versus macrophages/granulocytes isolated from cortex 24 hours after pMCAO. Light colored histograms represent cells stained with isotype control antibodies and filled histograms represent cells stained with antibodies for either IL-1β (A) or TNF-α (B).

From: Clausen BH, Lambertsen KL, Babcock AA, Holm TH, Dagnaes-Hansen F, Finsen B. Interleukin-1beta and tumor necrosis factor-alpha are expressed by different subsets of microglia and macrophages after ischemic stroke in mice.
J Neuroinflammation. 2008 Oct 23;5:46.

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Rat anti Mouse CD45 antibody, clone IBL-3/16 used for the detection of microglia using immunohistochemistry.
Image caption
Percentage of microgliosis (mean ± s.e.m) by area in (a) Tg APPsw/CD40 def. mice versus Tg APPsw mice and in (b) Tg PSAPP/CD40 def. mice versus Tg PSAPP mice at 22 to 24 months of age calculated by quantitative image analysis. Post hoc comparison between groups are indicated by the marked bars (* p < 0.05; ** p < 0.01). Representative photographs of brain area in (c) Tg APPsw, (e) Tg APPsw/CD40 def., (d) Tg PSAPP and (f) Tg PSAPP/CD40 def. mice stained with CD45 antibody (each bar represents 0.1 mm).

From: Laporte V, Ait-Ghezala G, Volmar CH, Mullan M. CD40 deficiency mitigates Alzheimer's disease pathology in transgenic mouse models.
J Neuroinflammation. 2006 Feb 24;3:3.

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Rat anti Mouse CD45 antibody, clone IBL-3/16 used for the demonstration of microglia busing immunohistochemistry.
Image caption
Chronic effects of celastrol on microgliosis in Tg PS1/APPsw mice. A) Representative photomicrographs (taken with a 20× and 60× objective providing a respective magnification of 200× and 600× respectively) depicting the presence of CD45 reactive microglia around Aß deposits in Tg PS1/APPsw treated with a placebo and celastrol. B) Histogram representing the burden of activated microglia (CD45 positive) in the cortex of Tg PS1/APPsw mice treated with placebo and celastrol pellets. Statistically significant difference in microgliosis burden (P < 0.03) was observed between placebo and celastrol treated mice. (* P < 0.05).

.From: Paris D, Ganey NJ, Laporte V, Patel NS, Beaulieu-Abdelahad D, Bachmeier C, March A, Ait-Ghezala G, Mullan MJ. Reduction of beta-amyloid pathology by celastrol in a transgenic mouse model of Alzheimer's disease.
J Neuroinflammation. 2010 Mar 8;7:17.

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Rat anti Mouse CD45 antibody, clone IBL-3/16 used for the detection of microglia by immunofluorescence.
Image caption:
Increased CD45+ microglial cells in hippocampus and cortex of the TgAPPsw mice with reduced GRK5. Representative IF results with anti-CD45 staining (green) in hippocampus (A-D) and cortex (F-I) of WT (A &F), heterozygote GRK5KO (B &G), TgAPPsw (C &H), and the double mice (D &I), respectively. Scale bar in panel I is for panels A-D and F-I: 100 µm. Panels E &J, examples of high magnification views of CD45+-microglial cells in hippocampus (E) and cortex (J) of the double mice that show details of activated microglial morphology. Scale bar in panel J is for panels E &J: 20 µm. Blue indicates reference DAPI staining of nuclei.

From: Li L, Liu J, Suo WZ. GRK5 deficiency exaggerates inflammatory changes in TgAPPsw mice.
J Neuroinflammation. 2008 Jun 3;5:24.

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Rat anti Mouse CD45 antibody, clone IBL-3/16 used for the detection of microglial cells by immunofluorescence.
Image caption:
Colocalization of activated microglial cells with Aβ plaques and IL-1β immunoreactivity in the TgAPPsw mice with reduced GRK5. Panels A-C, IF staining of Aβ+ plaques (A, red) and surrounding CD45+ microglial cells (B, green), as well as their merged view (C) in the double mice. Scale bar: 50 μm for panels A-C. Panels D-F, IF staining of Aβ+ plaques (D, red) and surrounding CD11b+ microglial cells (E, green), as well as their merged view (F) in the double mice. Scale bar: 50 μm for panels D-F. Panel G, an example of merged view for CD45 (green) and CD11b/c (clone OX42, red) co-staining of the microglial cells. The image showed that, at least in this particular experimental paradigm, the CD45 antibody stained more specifically for the microglial cell profiles; while the OX42 antibody, in addition to its positive staining of the microglial cells, also non-specifically decorated the plaques. Scale bar: 45 μm. Panel H, an example of merged view for Aβ+ plaques (red) and surrounding IL-1β immunoreactivity (green) in the double mice. Scale bar: 30 μm. Panel I, colocalization of CD45+ microglial cells (green) with IL-1β immunoreactivity (red) in the double mice. Scale bar: 30 μm. Blue indicates reference DAPI staining of nuclei.

From: Li et al. Journal of Neuroinflammation 2008 5:24.

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Rat anti Mouse CD45 antibody, clone IBL-3/16 used for the detection of leukocytes in mouse brain by immunofluorescence.
Image caption:
Cryosectioned brain samples from day 6 p.i. were stained for WNV envelope protein (green), CD45 (as a pan-leukocyte marker; red), and DAPI (blue) and utilized for laser scanning confocal microscopy (20X images are shown and are representative of n = 5 mice per group).

From: Wang P, Bai F, Zenewicz LA, Dai J, Gate D, et al. (2012) IL-22 Signaling Contributes to West Nile Encephalitis Pathogenesis.
PLoS ONE 7(8): e44153.

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Rat anti Mouse CD45 antibody, clone IBL-3/16 used for the detection of microglial cells by immunofluorescence.
Image caption:
Microglial distribution in minocycline-treated and untreated explants. The left panel shows microglial cells (stained with anti-CD45 antibody, green), whereas the right panel depicts both microglial cells and the location of retinal layers (revealed by nuclear staining with Hoechst, blue). A. Microglial cells in the control explants (CT) are frequently seen within the Outer Nuclear Layer (ONL). B. In minocycline-treated explants (MIN), microglial cells are more compact and do not penetrate the ONL. Representative images of at least three different explants per condition. INL, Inner Nuclear Layer; GCL, Ganglion Cell Layer. Scale bar, 50 μm.

From: Ferrer-Martín RM, Martín-Oliva D, Sierra-Martín A, Carrasco M-C, Martín-Estebané M, Calvente R, et al. (2015) Microglial Activation Promotes Cell Survival in Organotypic Cultures of Postnatal Mouse Retinal Explants.
PLoS ONE 10(8): e0135238.

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Rat anti Mouse CD45 antibody, clone IBL-3/16 used for the detection of microglial cells by immunofluorescence.
Image caption:
PARP-1 activity in untreated (A,B) and minocycline-treated (C,D) retinal explants. The left column shows the labeling with an antibody recognizing Poly-ADP-Ribose polymers (PAR, green) due to activity of the enzyme PARP-1. The central column exhibits microglial cells revealed by anti-CD45 antibody (red). Finally, pictures in the right column merge the two signals and that of Hoechst labeling of the nuclei. Note that minocycline-treatment (MIN) markedly reduces the PAR immunoreactivity of microglial cells in the control (CT) retinal explant. ONL, Outer Nuclear Layer; INL, Inner Nuclear Layer; GCL, Ganglion Nuclear Layer. Representative images of three retinal explants per treatment. Scale bar, 35 μm for A, 10 μm for B, 34 μm for C, and 9 μm for D.

From: Ferrer-Martín RM, Martín-Oliva D, Sierra-Martín A, Carrasco M-C, Martín-Estebané M, Calvente R, et al. (2015) Microglial Activation Promotes Cell Survival in Organotypic Cultures of Postnatal Mouse Retinal Explants.
PLoS ONE 10(8): e0135238.

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Rat anti Mouse CD45 antibody, clone IBL-3/16 used for the detection of microglial cells by immunofluorescence.
Image caption:
Effects of LPS treatment on retinal explants.
A: The release of TNF-α was significantly increased in explants treated with LPS (LPS, gray bar) with respect to untreated explants (CT, black bar). Bars represent mean values of 12 explants per condition ± SEM. *** indicates significant differences (P<0.001, Student's t-test). B: Incubation with LPS (LPS, gray bar) also greatly augmented the proportion of proliferating microglial cells (labeled with anti-CD11b and anti-Ki67) with respect to control explants (CT, black bar). Bars represent mean percentages of 5 explants per condition ± SEM; *** indicates significant differences (P<0.001, Student's t-test). C: The PAR immunostaining (green) in LPS explants was similar to that observed in the untreated explants. Pictures in C are representative of 3 explants per condition. ONL, Outer Nuclear Layer; INL, Inner Nuclear Layer; IPL, Inner Plexiform Layer; GCL, Ganglion Cell Layer. Scale bar, 30 μm.

From: Ferrer-Martín RM, Martín-Oliva D, Sierra-Martín A, Carrasco M-C, Martín-Estebané M, Calvente R, et al. (2015) Microglial Activation Promotes Cell Survival in Organotypic Cultures of Postnatal Mouse Retinal Explants.
PLoS ONE 10(8): e0135238.

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Rat anti Mouse CD45 antibody, clone IBL-3/16 used for the detection of microglial cells by immunofluorescence.
Image caption:
LPS treatment does not modify the distribution of microglial cell and cell viability.
A, B: Distribution of microglial cells in control (A, CT) and LPS-treated (B, LPS) retinal explants. Left panels display microglial cells labeled with anti-CD45 antibody, while right panels also show cell nuclei stained with Hoechst. The distribution of microglia in LPS-treated explants is similar to that of untreated explants. Representative images of 3 different explants per condition. Scale bar, 50 μm. C: Bar graph showing that LPS treatment has no significant effect on cell viability in retinal explants. Bars represent mean values ± SEM of 5 explants).

From: Ferrer-Martín RM, Martín-Oliva D, Sierra-Martín A, Carrasco M-C, Martín-Estebané M, Calvente R, et al. (2015) Microglial Activation Promotes Cell Survival in Organotypic Cultures of Postnatal Mouse Retinal Explants.
PLoS ONE 10(8): e0135238.

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CD45 Antibody | IBL-3/16 gallery image 14

Published customer image:
Rat anti Mouse CD45 antibody, clone IBL-3/16 used for the detection of microglial cells by immunofluorescence.
Image caption:
The effects of minocycline overcome these of LPS treatment. A: The cell viability in the explants treated with minocycline and LPS (MIN+LPS) is similar to that of explants treated with minocycline alone (MIN). Bars represent mean values ± SEM of 5 explants per condition. B, C: The distribution of microglial cells, as revealed by CD45 immunolabeling (green), is similar between explants treated with minocycline alone (MIN, B) and those treated with minocycline plus LPS (MIN+LPS, C). In addition to microglial cells, images in the right column also show the retinal layers revealed by the Hoechst staining of nuclei. Each image is representative of 3 explants. ONL, Outer Nuclear Layer; INL, Inner Nuclear Layer; GCL, Ganglion Cell Layer. Scale bar, 50 μm.

From: Ferrer-Martín RM, Martín-Oliva D, Sierra-Martín A, Carrasco M-C, Martín-Estebané M, Calvente R, et al. (2015) Microglial Activation Promotes Cell Survival in Organotypic Cultures of Postnatal Mouse Retinal Explants.
PLoS ONE 10(8): e0135238.

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  • Rat anti Mouse CD45
(Rated 0.0 out of 5 based on 0 customer reviews)
  • Product Type
    Monoclonal Antibody
  • Clone
    IBL-3/16
  • Isotype
    IgG1
1 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    MCA1388C *, F, IF, IPdatasheet pdfdatasheet pdf0.25 mg
    MCA1388
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
    -
    • Rat anti Mouse CD45 antibody, clone IBL-3/16 recognizes murine Receptor-type tyrosine-protein phosphatase C, also known as CD45, Leukocyte common antigen, T200 or Lymphocyte antigen 5 (Ly5). CD45 is a 1291 amino acid ~175kDa single pass type I transmembrane glycoprotein belonging to the protein -tyrosing phosphatase family. CD45 has two fibronectin type-III domains and two tyrosine protein phosphatase domains (UniProt:: P06800).

      Multiple isoforms are generated by alternative splicing with isoforms having differing deletions in the N-terminal region (Saga et al. 1987). Rat anti Mouse CD45 antibody, clone IBL-3/16 is expected to regognize all isoforms of murine CD45.

      Rat anti Mouse CD45 antibody, clone IBL-3/16 has been used sucessfully for the identification of CD45 in murine samples using Immunohistochemical (both cryo and FFPE), immunofluorescence and western blotting techniques (Kondo et al. 2011, Cuadros et al. 2006).
    • 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
      Flow Cytometry1/501/100
      Immunofluorescence
      Immunohistology - Paraffin
      Immunohistology - Resin
      Immunoprecipitation
      Western Blotting
      Immunohistology - Frozen(1)1/501/100
      (1)
      The epitope recognised by this antibody is reported to be sensitive to formaldehyde fixation and tissue processing. Bio-Rad recommends the use of acetone fixation for frozen sections.

    • 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
    • Recommended Protocol
    • Flow Cytometry
      Use 10ul of the suggested working dilution to label 106 cells in 100ul.
    • ELISA
    • Immunohistology
    • Histology Positive Control Tissue
    • Immunofluorescence
    • Western Blotting
    • Instructions For Use

    Additional CD45 Antibody Formats

    Formats Clone Applications Sizes available
    CD45 Antibody : Purified IBL-3/16 C *, F, IF, IP 0.25 mg
    • Copyright © 2016 Bio-Rad

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      Recommended Negative Isotype Control

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        Useful Reagents

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              References

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