The PhosphoPlus® Rb (Ser780, Ser807/811) Antibody Kit provides reagents and protocols to investigate cell cycle progression within cells. The kit contains a total Rb antibody, two distinct phospho-Rb antibodies, and Rb control proteins along with secondary antibodies and reagents to perform up to 10 western blots.
Phospho-Rb (Ser780) (D59B7) Rabbit mAb and Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb detect endogenous levels of Rb only when phosphorylated at Ser780 and Ser807/811, respectively. Rb (4H1) Mouse mAb detects endogenous levels of total Rb protein. The antibody does not cross-react with the Rb homologues p107 or p130, or with other proteins.
Phospho-Rb (Ser780) (D59B7) Rabbit mAb is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser780 of human Rb protein. Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser807/811 of human Rb protein. Rb (4H1) monoclonal antibody is produced by immunizing animals with Rb-C Fusion Protein #6022, which contains residues 701-928 of human Rb .
The retinoblastoma tumor suppressor protein Rb regulates cell proliferation by controlling progression through the restriction point within the G1-phase of the cell cycle (1). Rb has three functionally distinct binding domains and interacts with critical regulatory proteins including the E2F family of transcription factors, c-Abl tyrosine kinase, and proteins with a conserved LXCXE motif (2-4). Cell cycle-dependent phosphorylation by a CDK inhibits Rb target binding and allows cell cycle progression (5). Rb inactivation and subsequent cell cycle progression likely requires an initial phosphorylation by cyclin D-CDK4/6 followed by cyclin E-CDK2 phosphorylation (6). Specificity of different CDK/cyclin complexes has been observed in vitro (6-8) and cyclin D1 is required for Ser780 phosphorylation in vivo (9).
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