Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

Phospho-SirT1 (Ser47) Antibody #2314

Applications Reactivity Sensitivity MW (kDa) Source
W IP IF-IC F H Endogenous 120 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-SirT1 (Ser47) Antibody detects endogenous levels of SirT1 protein only when phosphoryated at serine 47. The antibody does not cross-react with other sirtuin proteins.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser47 of human SirT1. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of 293 cell lysates treated or untreated with λ phosphatase, using Phospho-SirT1 (Ser47) Antibody #2314 (upper) or SirT1 Antibody #2310 (lower).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of K-562 cells, λ phosphatase treated (blue) or untreated (green), using Phospho-SirT1 (Ser47) Antibody compared to a nonspecific negative control antibody (red).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HT-1080 cells, untreated (left) and phosphatase-treated (right), using Phospho-SirT1 (Ser47) Antibody (green). Actin filaments have been labeled using DY-554 phalloidin (red).


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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