Anti-Rad21 (S. pombe) antibody, rabbit serum

Description

Rad21 protein (628 aa, 67.8 kDa) is a cleavable component of the cohesin complex, involved in chromosome cohesion during cell cycle. The cohesin complex is required for the cohesion of sister chromatids after DNA replication. The cohesin complex apparently forms a large proteinaceous ring within which sister chromatids can be trapped. At metaphase-anaphase transition, this protein is cleaved by Cut1 and dissociates from chromatin, allowing sister chromatids to segregate. Also involved in the DNA double-strand-break (DSB) repair system hyperphosphorylated during S and G2 phases. Proteolytic cleavage of a fraction of hyperphosphorylated form at the onset of anaphase may be essential for the proper progression of anaphase and sister chromatid separation. Belongs to the rad21 family, conserved to mammals.

Applications

Western blot 1/1,000~1/4,000 dilution (Ref.1 & 2). Not tested for other applications

Specification

Immunogen: Recombinant full-size S. pombe Rad21 protein (His-tagged) expressed in E. coli

Specificity: Reacts with S. pombe Rad21 protein. Not tested with other species

Form: Rabbit antiserum with 0.05 % sodium azide

Storage: Ship at 4℃ and long term storage at -20℃ or below

Data Link: S. pombe Gene DB: rad21

References: This antibody was used in the following references.

1. Nagao K, Adachi Y, Yanagida M. Separase-mediated cleavage required
2. Adach Y et al. Cut1/separase-dependent roles of multiplefor DNA repair. Nature. (2004) 430:1044-8. pubmed/15329725
   phosphorylation of fission yeast cohesion subunit Rad21 in post-replicative damage repair and mitosis. Cell Cycle. (2008) 15:765-76. pubmed/18239448

Figure: Identification of Rad21 protein in crude extract of S. pombe with anti-Rad21 antiserum.

WT cells and rad21-9xPK cells grown in YS5S to 1×10⁷, collected, washed and broken by glass beads. Twenty ug proteins per lane were run on SDS-PAGE. First antibody at indicated dilutions in PBST plus 2.5% skim milk was incubated at 4°C over-night. Second antibody at 1/10,000 of anti-rabbit HRP in PBST was incubated at room temperature for 1 h. Detected by ECL (X-ray film). Rad21 protein migrated at the position of ~110 kDa in SDS-PAGE although the molecular mass is 67.8 kDa (Ref. 2). Multiple bands of Rad21 reflect multiple phosphorylation states.