Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

SET8 (C18B7) Rabbit mAb #2996

Applications Reactivity Sensitivity MW (kDa) Isotype
W IF-IC H M R Mk (B) (Pg) (Hr) Endogenous 43, 50 Rabbit IgG

Applications Key:  W=Western Blotting  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  B=Bovine  Pg=Pig  Hr=Horse
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

SET8 (C18B7) Rabbit mAb detects endogenous levels of total SET8 protein (both isoforms).

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to human SET8 protein. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from HCT116 and RAW cells using SET8 (C18B7) Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with 0.5% Triton X-100 for 10 minutes prior to fixation according to Tardat et al. (2007) J. Cell Biol. 179, 1413-26 (right), using SET8 (C18B7) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).

Background

SET domain-containing lysine methyltransferase 8 (SET8), also known as PR/SET domain-containing protein 7 (PR/SET7), is a member of a family of histone lysine methyltransferases, each of which contains a conserved catalytic SET domain originally identified in Drosophila Su[var]3-9, Enhancer of zeste, and Trithorax proteins (1-3). SET8 is a single-subunit enzyme that mono-methylates histone H4 on Lys20, preferably on nucleosomal substrates (1-3). SET8 protein levels and Histone H4 Lys20 methylation are cell cycle regulated, both increasing in S phase and peaking at G2/M phase (4,5). SET8 interacts with the PCNA protein, associates with sites of active DNA synthesis and is required for DNA replication and genome stability during S phase (5-7). Inhibition of SET8 using shRNA results in arrest of replication forks, induction of double-stranded DNA breaks and a Chk1-mediated cell-cycle arrest in S and G2/M phases of the cell cycle (6,7). Furthermore, SET8 methylates p53 on Lys382, down regulating the pro-apoptotic and checkpoint activation functions of p53 (8). In response to DNA damage, SET8 expression levels decrease, allowing p53 to activate checkpoints and/or apoptosis (8). Both the methylation of histone H4 Lys20 and p53 appear to be important for the functions of SET8 in S phase.

  1. Fang, J. et al. (2002) Curr Biol 12, 1086-99.
  2. Xiao, B. et al. (2005) Genes Dev 19, 1444-54.
  3. Couture, J.F. et al. (2005) Genes Dev 19, 1455-65.
  4. Rice, J.C. et al. (2002) Genes Dev 16, 2225-30.
  5. Huen, M.S. et al. (2008) J Biol Chem 283, 11073-7.
  6. Tardat, M. et al. (2007) J Cell Biol 179, 1413-26.
  7. Jørgensen, S. et al. (2007) J Cell Biol 179, 1337-45.
  8. Shi, X. et al. (2007) Mol Cell 27, 636-46.

Application References

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Companion Products

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

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