Anti-BrdU antibody [IIB5] (ab8152) is a mouse monoclonal antibody detecting BrdU in Flow Cytometry, IHC-P, IHC-Fr, ICC/IF.
- Over 80 publications
- Trusted since 2002
pH: 7.3
Preservative: 0.09% Sodium azide
Constituents: PBS, 1% BSA
Flow Cyt | IHC-FoFr | IHC-P | ICC/IF | IHC-Fr | |
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Chemical | Tested | Expected | Tested | Tested | Expected |
Species | Dilution info | Notes |
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Species Chemical | Dilution info 1/100.00000 - 1/200.00000 | Notes ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
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Species Chemical | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
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Species Chemical | Dilution info 1/5.00000 - 1/10.00000 | Notes Enzymatic antigen retrieval with proteases can also be used. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
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Species Chemical | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Chemical | Dilution info 1/5.00000 - 1/10.00000 | Notes Dilute antibody in 0.15 M phosphate buffered saline with 1% BSA and 1% Na-azide. |
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BUdr, Bromodeoxyuridine
Anti-BrdU antibody [IIB5] (ab8152) is a mouse monoclonal antibody detecting BrdU in Flow Cytometry, IHC-P, IHC-Fr, ICC/IF.
- Over 80 publications
- Trusted since 2002
pH: 7.3
Preservative: 0.09% Sodium azide
Constituents: PBS, 1% BSA
BrdU is a thymidine analogue and when offered to proliferating cells it is incroporated into reduplicating cells. The antibody is specific for DNA in which BrdU has been incorporated. In immunoassays this antibody reacts strongly with free or carrier-protein coupled BrdU but not with other nucleosides. In immuncytochemistry the antibody only recognizes BrdU in denaturated (single stranded) DNA. The BrdU antibody is 100% crossreactive with Iodo-Deoxy-Uridine (IrdU). Therfore, IdU instead of BrdU can be used in studies.
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This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
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Full details and terms and conditions can be found here:
Terms & Conditions.
Immunohistochemical analysis of rat brain tissue, staining BrdU with ab8152.
Tissue was fixed with paraformaldehyde and blocked with 5% serum for 1 hour at 25°C; antigen retrieval was by heat mediation in citrate buffer (pH 6). Samples were incubated with primary antibody (1/20 in diluent) for 18 hours at 25°C. A Cy3®-conjugated donkey anti-mouse polyclonal IgG was used as the secondary antibody.
ab8152 (1/100) staining BrdU in HeLa cells (green). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X100/ PBS and counterstained with DAPI in order to highlight the nucleus (red). For further esperimental details please see Abreview.
Cells were pulse labeled with 10 μM BrdU for 30 min, rinsed twice in prewarmed PBS, and chased in prewarmed culture medium, supplemented with 5 mM deoxythymidine. Incorporated BrdU was detected after ethanol fixation of the cells, which were than rinsed once in PBS and resuspended in 2 ml of 0.4 mg/ml pepsin in 0.1 N HCl. After 30 min at room temperature cells were pelleted, resuspended in 2 N HCl, and incubated for another 30 min at 37°C. Cells were rinsed in 0.1 M borate buffer, pH 8.5, and PBS/BSA (1 mg/ml BSA in PBS). Appropriately diluted mouse anti-BrdU antibody (clone IIB5) was added to the cell pellet, resuspended in 100 micro liters PBS/BSA. After incubation for 1 h at room temperature, the cells were rinsed twice in PBS/BSA. For visualization, FITC-conjugated Fab2 fragments of rabbit anti-mouse Ig antibody were added in a 1/10 dilution. After incubation for 45 min at room temperature samples were rinsed twice in PBS/BSA and the cells were finally resuspended in 0.5 ml cold PBS supplemented with 100 microgram/ml RNAse and 20 μg/mL propidium iodide. The samples were allowed to stand for 15 min on ice in the dark before flow cytometric analysis. In the negative control the primary antibody was omitted.
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