Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

SirT1 (D1D7) Rabbit mAb #9475

Applications Reactivity Sensitivity MW (kDa) Isotype
W IF-IC H M R Mk (C) (B) (Pg) (Hr) Endogenous 120 Rabbit IgG

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

SirT1 (D1D7) Rabbit mAb recognizes endogenous levels of total SirT1 protein. This antibody does not cross-react with other sirtuin proteins.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Phe297 of human SirT1 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using SirT1 (D1D7) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from SirT1 wild-type (WT) and knockout (KO) mouse embryonic fibroblasts (MEF) using SirT1 (D1D7) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). WT and KO MEF were kindly provided by Wenyi Wei, Harvard Medical School.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa (positive; left), WT MEF (positive; middle), and SirT1 KO MEF (right) cells using SirT1 (D1D7) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). WT and KO MEF were kindly provided by Wenyi Wei, Harvard Medical School.


Background

The Silent Information Regulator (SIR2) family of genes is a highly conserved group of genes that encode nicotinamide adenine dinucleotide (NAD)-dependent protein deacetylases, also known as class III histone deacetylases. The first discovered and best characterized of these genes is Saccharomyces cerevisiae SIR2, which is involved in silencing of mating type loci, telomere maintenance, DNA damage response, and cell aging (1). SirT1, the mammalian ortholog of Sir2, is a nuclear protein implicated in the regulation of many cellular processes, including apoptosis, cellular senescence, endocrine signaling, glucose homeostasis, aging, and longevity. Targets of SirT1 include acetylated p53 (2,3), p300 (4), Ku70 (5), forkhead (FoxO) transcription factors (5,6), PPARγ (7), and the PPARγ coactivator-1α (PGC-1α) protein (8). Deacetylation of p53 and FoxO transcription factors represses apoptosis and increases cell survival (2,3,5,6). Deacetylation of PPARγ and PGC-1α regulates the gluconeogenic/glycolytic pathways in the liver and fat mobilization in white adipocytes in response to fasting (7,8). SirT1 deacetylase activity is inhibited by nicotinamide and activated by resveratrol. In addition, SirT1 activity may be regulated by phosphorylation, as it is phosphorylated at Ser27 and Ser47 in vivo; however, the function of these phosphorylation sites has not yet been determined (9).

  1. Guarente, L. (1999) Nat. Genet. 23, 281-285.
  2. Vaziri, H. et al. (2001) Cell 107, 149-159.
  3. Luo, J. et al. (2001) Cell 107, 137-148.
  4. Bouras, T. et al. (2005) J. Biol. Chem. 280, 10264-10276.
  5. Brunet, A. et al. (2004) Science 303, 2011-2015.
  6. Motta, M.C. et al. (2004) Cell 116, 551-563.
  7. Picard, F. et al. (2004) Nature 429, 771-776.
  8. Rodgers, J.T. et al. (2005) Nature 434, 113-118.
  9. Beausoleil, S.A. et al. (2004) Proc. Natl. Acad. Sci. USA 101, 12130-12135.

Application References

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

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