Nonphosphorylated Rb-C Fusion Protein (5 µg/ml): Rb-C is expressed as a recombinant fusion protein of Rb residues 701–928 and maltose binding protein, serves as a negative control. Supplied in SDS Sample Buffer. Phosphorylated Rb-C Fusion Protein (5 µg/ml): Prepared by in vitro kinase reaction with cdc2, serves as a positive control. Supplied in SDS Sample Buffer.
Directions for Use
These lysates are useful controls for total and Phospho-Rb antibodies. However, this truncated Rb recombinant protein is not recognized by Phospho-Rb (Ser608) Antibody #2181 or Rb (D20) Rabbit mAb #9313.
Background
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).
Storage: Supplied in SDS Sample Buffer:
62.5 mM Tris-HCl (pH 6.8 at 25°C), 2% w/v SDS,
10% glycerol, 50 mM DTT, 0.01% w/v bromophenol
blue. Store at –20°C
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