Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

RAD21 (D213) Antibody #4321

Applications Reactivity Sensitivity MW (kDa) Source
W IP IF-IC H M R Mk (C) (B) (Pg) (Hr) Endogenous 130 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  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

RAD21 (D213) Antibody detects endogenous levels of total RAD21 protein.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the human RAD21 protein. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using RAD21 (D213) Antibody.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using RAD21 (D213) Antibody (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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