Product Pathways - Cell Cycle / Checkpoint
BrdU (Bu20a) Mouse mAb #5292
| Applications | Reactivity | Isotype |
|---|---|---|
| IHC-P IF-IC F | All | Mouse IgG1 |
Applications Key:
IHC-P=Immunohistochemistry (Paraffin)
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key: All=All species expected
Protocols
- 5292:
- Flow, IHC / Paraffin, Immunofluorescence
Specificity / Sensitivity
BrdU (Bu20a) Mouse mAb detects BrdU when incorporated into single stranded DNA. DNA must be denatured for the epitope to be exposed and recognized by the antibody.
Source / Purification
Monclonal antibodies are produced by immunizing animals with BrdU conjugated to BSA.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded HeLa cells, without (left) and with (right) BrdU incorporation, using BrdU (Bu20a) Mouse mAb.
Flow Cytometry
Flow cytometric analysis of Jurkat cells incorporated with BrdU (30 minutes), using BrdU (Bu20a) Mouse mAb versus propidium iodide (DNA content).
Flow Cytometry
Flow cytometric analysis of Jurkat cells, untreated (blue) or BrdU incorporated for 30 minutes (green), using BrdU (Bu20a) Mouse mAb.
IF-IC
Confocal immunofluorescent analysis of HeLa cells using BrdU (Bu20a) Mouse mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Background
Halogenated nucleotides such as the pyrimidine analog bromodeoxyuridine (BrdU) are useful for labeling nascent DNA in living cells and tissues. BrdU becomes incorporated into replicating DNA in place of thymidine and subsequent immunodetection of BrdU using specific monoclonal antibodies allows labeling of cells in S phase of the cell cycle. After pulse-labeling cells or tissues with bromodeoxyuridine, BrdU (Bu20a) Mouse mAb can be used to detect BrdU incorporated into single stranded DNA. Please see our detailed protocol for information regarding the labeling procedure as well as denaturation of double stranded DNA for various immunodetection applications (1-4).
- Darzynkiewicz, Z. and Juan, G. (2001) Curr Protoc Cytom Chapter 7, Unit 7.7.
- Leif, R.C. et al. (2004) Cytometry A 58, 45-52.
- Staszkiewicz, J. et al. (2009) Biochem Biophys Res Commun 378, 539-44.
- Rothaeusler, K. and Baumgarth, N. (2007) Curr Protoc Cytom Chapter 7, Unit7.31.
Application References
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
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This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
