Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

Phospho-SMC1 (Ser360) Antibody #4029

Applications Reactivity Sensitivity MW (kDa) Source
W IF-IC H M R (Mk) (C) (X) (B) (Sc) Endogenous 145 Rabbit

Applications Key:  W=Western Blotting  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  C=Chicken  X=Xenopus  B=Bovine  Sc=S. cerevisiae
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-SMC1 (Ser360) Antibody detects endogenous levels of SMC1 protein only when phosphorylated on Ser360. This antibody does not cross-react with other SMC proteins.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to Ser360 of the human SMC1 protein. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells, untreated or UV-treated (100 J/m2 followed by 4 hour recovery) using Phospho-SMC1 (Ser360) Antibody (upper) or SMC1 Antibody #4802 (lower). Antibody phospho-specificity was determined by treating cell extracts with λ phosphatase.

IF-IC

IF-IC

Confocal immunfluorescent analysis of HeLa cells, untreated (left) and UV-treated (right), using Phospho-SMC1 (Ser360) Antibody (green). Actin filaments have been labeled with DY-554 phalloidin (red).

Background

Structural maintenance of chromosomes 1 (SMC1) protein is a chromosomal protein member of the cohesin complex that enables sister chromatid cohesion and plays a role in DNA repair (1,2). ATM/NBS1-dependent phosphorylation of SMC1 occurs at Ser957 and Ser966 in response to ionizing radiation (IR) as part of the intra-S-phase DNA damage checkpoint (3). SMC1 phosphorylation is ATM-independent in cells subjected to other forms of DNA damage, including UV light and hydroxyurea treatment (4). While phosphorylation of SMC1 is required for activation of the IR-induced intra-S-phase checkpoint, the precise mechanism is not well understood and may involve a conformational change that affects SMC1-SMC3 interaction (3).

The serine residue at 360 of SMC1 is phosphorylated in an ATM/ATR-dependent manner in response to DNA damage (5,6). Phospho-SMC1 (Ser360) Antibody is directed at a site that was identified at Cell Signaling Technology (CST) using PhosphoScan®, CST's LC-MS/MS platform for modification site discovery. Phosphorylation at Ser360 was discovered using an ATM/ATR substrate antibody and was shown to be induced by UV treatment. Please visit PhosphoSitePlus®, CST's modification site knowledgebase, at www.phosphosite.org for more information.

  1. Michaelis, C. et al. (1997) Cell 91, 35-45.
  2. Sjögren, C. and Nasmyth, K. (2001) Curr Biol 11, 991-5.
  3. Yazdi, P.T. et al. (2002) Genes Dev 16, 571-82.
  4. Kim, S.T. et al. (2002) Genes Dev 16, 560-70.
  5. Stokes, M.P. et al. (2007) Proc Natl Acad Sci U S A 104, 19855-60.
  6. Matsuoka, S. et al. (2007) Science 316, 1160-6.

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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