Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

MacroH2A1.2 Antibody #4827

Applications Reactivity Sensitivity MW (kDa) Source
W IF-IC H M R Mk (C) (B) Endogenous 40 Rabbit

Applications Key:  W=Western Blotting  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat  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

MacroH2A1.2 Antibody detects endogenous levels of the core histone MacroH2A1.2 protein (MacroH2A1, isoform 2). The antibody does not cross-react with MacroH2A1.1 (MacroH2A1, isoform 1), MacroH2A2 or histone H2A.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the human MacroH2A1.2 protein (MacroH2A1, isoform 2). Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, H-4-II-E and COS cells using MacroH2A1.2 Antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, either untransfected or transfected with expression constructs for MacroH2A1.1 or MacroH2A1.2, using MacroH2A1.2 Antibody (upper) and MacroH2A1.1 Antibody #4160 (lower).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using MacroH2A1.2 Antibody (green). Actin filaments were labeled using DY-554 phalloidin (red).


Background

Histone macroH2A1 and macroH2A2 comprise a family of variant histone H2A proteins. MacroH2A1 exists as two distinct isoforms due to alternative splicing of a single gene; macroH2A1.1 levels accumulate throughout differentiation and development while macroH2A1.2 shows a constant level of expression (1). MacroH2A1 and macroH2A2 are encoded by completely distinct genes located on separate chromosomes (2,3). Both macroH2A1 and macroH2A2 proteins contain an amino-terminal histone-like region with 64% sequence identity to canonical histone H2A, in addition to a carboxy-terminal “macro” domain (1-3). MacroH2A1 and macroH2A2 are enriched in facultative heterochromatin, including inactivated X chromosomes in mammalian females and senescence-associated heterochromatin foci (2-5). Both act to repress gene transcription by inhibiting the binding of transcription factors to chromatin, the acetylation of histones by p300, and the chromatin-remodeling activities of SWI/SNF and ACF (6,7). The macro domain of macroH2A1.1 binds to ADP-ribose and functions to recruit macroH2A1.1 to activated PARP at sites of DNA damage, where it mediates chromatin rearrangements to locally regulate the DNA damage response (8). MacroH2A1.2 and macroH2A2 do not bind poly-ADP-ribose and are not recruited to sites of activated PARP (8).

  1. Pehrson, J.R. et al. (1997) J Cell Biochem 65, 107-13.
  2. Chadwick, B.P. and Willard, H.F. (2001) Hum Mol Genet 10, 1101-13.
  3. Costanzi, C. and Pehrson, J.R. (2001) J Biol Chem 276, 21776-84.
  4. Costanzi, C. and Pehrson, J.R. (1998) Nature 393, 599-601.
  5. Zhang, R. et al. (2005) Dev Cell 8, 19-30.
  6. Angelov, D. et al. (2003) Mol Cell 11, 1033-41.
  7. Doyen, C.M. et al. (2006) Mol Cell Biol 26, 1156-64.
  8. Timinszky, G. et al. (2009) Nat Struct Mol Biol 16, 923-9.

Application References

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Companion Products

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.

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