Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

ASF1A (C6E10) Rabbit mAb #2990

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IHC-P IF-IC H M Mk (C) (B) Endogenous 20 kDa Rabbit IgG

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

Protocols

Specificity / Sensitivity

ASF1A (C6E10) Rabbit mAb detects endogenous levels of total ASF1A protein. The antibody does not cross-react with ASF1B protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the carboxy terminus of the human ASF1A protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using ASF1A (C6E10) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human melanoma using ASF1A (C6E10) Rabbit mAb in the presence of control peptide (left) or antigen specific peptide (right).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using ASF1A (C6E10) Rabbit mAb (green). Actin filaments have been labeled with DY-554 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. Silljé, 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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For Research Use Only. Not For Use In Diagnostic Procedures.

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