Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

SMC3 (D47B5) Rabbit mAb #5696

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC H M R Mk (C) (X) (Z) (B) Endogenous 140 Rabbit IgG

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

SMC3 (D47B5) Rabbit mAb recognizes endogenous levels of total SMC protein. In immunofluorescent analysis, this antibody shows nuclear staining as expected; however, it also shows weak cytoplasmic staining that is presumed to be background.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg132 of human SMC3 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using SMC3 (D47B5) Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using SMC3 (D47B5) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).

Background

The cohesin complex consists of a heterodimer between SMC1 (SMC1A or B) and SMC3, bound by additional RAD21 and STAG proteins (STAG1, 2, or 3) (1,2). These proteins form a ring-like structure that mediates the cohesion of two sister chromatids after DNA replication in S phase (1,2). RAD21 and STAG2 are phosphorylated by Polo-like kinase (PLK) during prophase, which leads to the dissociation of cohesin complexes from the chromosome arms; however, cohesin remains bound to centromeres until anaphase (3,4). RAD21 is cleaved by separin/ESPL1 in anaphase, which leads to dissociation of the remaining cohesin from centromeres, enabling sister chromatids to segregate during mitosis (5). RAD21 is also cleaved by caspase-3 and caspase-7 during apoptosis, resulting in a 64 kDa carboxy-terminal cleavage product that translocates to the cytoplasm and may help to trigger apoptosis (6,7). In addition to mediating cohesion of sister chromatids, the cohesin complex plays important roles in gene regulation and DNA repair, as SMC1 and SMC3 are both phosphorylated by ATM and ATR kinases upon DNA damage (1,2).

  1. Peters, J.M. et al. (2008) Genes Dev 22, 3089-114.
  2. Barbero, J.L. (2009) Cell Mol Life Sci 66, 2025-35.
  3. Hoque, M.T. and Ishikawa, F. (2001) J Biol Chem 276, 5059-67.
  4. Hauf, S. et al. (2005) PLoS Biol 3, e69.
  5. Hauf, S. et al. (2001) Science 293, 1320-3.
  6. Pati, D. et al. (2002) Mol Cell Biol 22, 8267-77.
  7. Chen, F. et al. (2002) J Biol Chem 277, 16775-81.

Application References

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Companion Products

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

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