Cell Signaling Technology
XP Monoclonal Antibody

Product Pathways - Chromatin Regulation / Epigenetics

CHAF1A (D77D5) XP® Rabbit mAb #5480

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC H Mk Endogenous 145 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

CHAF1A (D77D5) XP® Rabbit mAb detects endogenous levels of total CHAF1A protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro303 of human CHAF1A protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, Jurkat, and COS-7 cell lines using CHAF1A (D77D5) XP® Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using CHAF1A (D77D5) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).

Background

Chromatin assembly factor 1 (CAF-1) is a histone H3/H4 chaperone complex that functions in de novo assembly of nucleosomes during DNA replication and nucleotide excision repair (1). Nucleosome assembly is a two-step process, involving initial deposition of a histone H3/H4 tetramer onto DNA, followed by the deposition of a pair of histone H2A/H2B dimers (1). CAF-1 interacts with PCNA and localizes to DNA replication and DNA repair foci, where it functions to assemble newly synthesized histone H3/H4 tetramers onto replicating DNA (2-6). Assembly of histone H2A/H2B dimers requires additional assembly factors. The CAF-1 complex consists of three proteins: CHAF1A (p150), CHAF1B (p60) and RBAP48 (p48 or RBBP4). CHAF1A and CHAF1B proteins are specific for the CAF-1 complex, while RBAP48 is a component of multiple chromatin modifying complexes (1). CHAF1A and CHAF1B expression levels correlate with cellular proliferation and both proteins are significantly down-regulated in quiescent cells (7).

  1. Adams, C.R. and Kamakaka, R.T. (1999) Curr Opin Genet Dev 9, 185-90.
  2. Smith, S. and Stillman, B. (1989) Cell 58, 15-25.
  3. Smith, S. and Stillman, B. (1991) EMBO J 10, 971-80.
  4. Shibahara, K. and Stillman, B. (1999) Cell 96, 575-85.
  5. Moggs, J.G. et al. (2000) Mol Cell Biol 20, 1206-18.
  6. Gaillard, P.H. et al. (1996) Cell 86, 887-96.
  7. Polo, S.E. et al. (2004) Cancer Res 64, 2371-81.

Application References

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Companion Products

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

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