Phospho-RB-S811 Rabbit pAb for WB, ELISA - P06400

Core Product Specifications and Parameters

Biological Background: RB Function and Localization

• RB is a tumor suppressor protein that critically regulates the G1/S transition of the cell cycle by controlling E2F-dependent transcription. Related references: PMID:10499802
• In its hypophosphorylated form, RB binds E2F transcription factors, physically blocking their transactivation domain and recruiting chromatin-modifying enzymes to repress gene transcription. Related references: PMID:10499802
• Cyclin/CDK-mediated phosphorylation of RB induces its dissociation from E2Fs, enabling transcription of S-phase genes and cell cycle progression; phosphorylation by CDK3/cyclin-C specifically promotes the G0-G1 transition. Related references: PMID:10499802 PMID:15084261
• RB also contributes to heterochromatin formation by stabilizing histone methylation and recruiting histone methyltransferases such as SUV39H1, thereby maintaining constitutive heterochromatin structure.
• Subcellularly, RB localizes primarily to the nucleus but can also be detected in the cytoplasm.
• Post-translational modifications including phosphorylation, acetylation, and methylation regulate RB’s activity; viral oncoproteins (e.g., SV40 large T antigen, HPV E7) can disrupt RB-E2F complexes, highlighting its role in host-virus interactions.
• Tissue expression includes the retina and foreskin keratinocytes (at protein level). Related references: PMID:20940255

Experimental Guidance and Technical Tips

• For Western blot analysis, include phosphatase inhibitors in lysis buffers to preserve phosphorylation of RB at Ser811.
• Validate antibody specificity by peptide competition assay using the phosphorylated immunogen peptide or by phosphatase treatment of cell lysates.
• Select cell lines or tissue samples with known RB phosphorylation at Ser811 (e.g., cycling cancer cell lines) as positive controls.
• For ELISA applications, titrate the antibody to determine optimal coating and detection conditions, performing standard curve assays with phospho-peptide and non-phospho controls to confirm specificity.

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