Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

BrdU (Bu20a) Mouse mAb #5292

Applications Reactivity Sensitivity Isotype
IHC-P IF-IC F All Endogenous Mouse IgG1

Applications Key:  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key: All=All species expected
Species cross-reactivity is determined by western blot.

Protocols

* Product-specific protocol.

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

Monoclonal antibody is produced by immunizing animals with BrdU conjugated to BSA.

IHC-P (paraffin)

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 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 Cytometry

Flow cytometric analysis of Jurkat cells, untreated (blue) or BrdU incorporated for 30 minutes (green), using BrdU (Bu20a) Mouse mAb.


IF-IC

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 and denaturation of double stranded DNA for various immunodetection applications (1-4).

  1. Darzynkiewicz, Z. and Juan, G. (2001) Curr Protoc Cytom Chapter 7, Unit 7.7.
  2. Leif, R.C. et al. (2004) Cytometry A 58, 45-52.
  3. Staszkiewicz, J. et al. (2009) Biochem Biophys Res Commun 378, 539-44.
  4. Rothaeusler, K. and Baumgarth, N. (2007) Curr Protoc Cytom Chapter 7, Unit7.31.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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