Cell Signaling Technology

Product Pathways - Chromatin Regulation

ASF1B (C70E2) Rabbit mAb #2902

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC H Mk Endogenous 19 Rabbit

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.

Specificity / Sensitivity

ASF1B (C70E2) Rabbit mAb detects endogenous levels of total ASF1B protein. The antibody does not cross-react with ASF1A protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide (KLH-coupled) corresponding the the carboxy-terminus of the human ASF1B protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and HCT116 cells using ASF1B (C70E2) Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using ASF1B (C70E2) Rabbit mAb (green). Actin filaments have been labeled with DY554 phalloidin (red).

Background

ASF1 was first identified in S. cerevisiae based on its ability to de-repress transcriptional silencing when overexpressed (1). While only one gene exists in yeast and Drosophila, mammalian cells contain the two highly homologous ASF1A and ASF1B genes (2). ASF1A and ASF1B function as histone chaperones, delivering histone H3/H4 dimers to CAF-1 or HIRA histone deposition complexes to facilitate replication-coupled and replication-independent nucleosome assembly on DNA (2-5). Both ASF1A and ASF1B bind to CAF-1, but only ASF1A binds to HIRA (5). In addition to playing a role in DNA replication and gene silencing, ASF1 functions in DNA damage repair, genome stability and cellular senescence. Deletion of ASF1 in yeast and Drosophila confers sensitivity to various DNA damaging agents and inhibitors of DNA replication, increases genomic instability and sister chromatid exchange, and activates the DNA damage checkpoint (6-8). Depletion of both ASF1A and ASF1B in mammalian cells results in the accumulation of cells in S phase, increased phosphorylation of H2A.X, centrosome amplification and apoptosis (9,10). ASF1A is required for the formation of senescence-associated heterochromatin foci (SAHF), with overexpression of ASF1A inducing senescence in primary cells (4). Both ASF1A and ASF1B are phosphorylated in S phase by the Tousled-like kinases TLK1 and TLK2, and are dephosphorylated when TLK1 and TLK2 are inactivated by Chk1 kinase in response to replicative stress (11,12). The function of ASF1 phosphorylation is not yet understood.

  1. Singer, M.S. et al. (1998) Genetics 150, 613-632.
  2. Mousson, F. et al. (2007) Chromosoma 116, 79-93.
  3. Tang, Y. et al. (2006) Nat. Struct. Mol. Biol. 13, 921-929.
  4. Zhang, R. et al. (2005) Dev. Cell. 8, 19-30.
  5. Daganzo, S.M. et al. (2003) Curr. Biol. 13, 2148-2158.
  6. Ramey, C.J. et al. (2004) Mol. Cell. Biol. 24, 10313-10327.
  7. Prado, F. et al. (2004) EMBO Rep. 5, 497-502.
  8. Tyler, J.K. et al. (1999) Nature 402, 555-560.
  9. Sanematsu, F. et al. (2006) J. Biol. Chem. 281, 13817-13827.
  10. Groth, A. et al. (2005) Mol. Cell. 17, 301-311.
  11. Sillje, H.H. and Nigg, E.A. (2001) Curr. Biol. 11, 1068-1073.
  12. Carrera, P. et al. (2003) Genes Dev. 17, 2578-2590.

Application References

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This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.

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