Product Pathways - Cell Cycle / Checkpoint
PhosphoPlus® Rb (Ser780, Ser807/811) Antibody Kit #9306
|9306S||1 Kit (10 western blots)||---||In Stock||---|
|9306||carrier free and custom formulation / quantity||email request|
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|Kit Includes||Quantity||Applications||Reactivity||Homology†||Mol. Wt.||Isotype|
|Phospho-Rb (Ser780) Antibody #9307||100 µl||W, IP, E-P||H, R, Mk||110 kDa||Rabbit|
|Phospho-Rb (Ser807/811) Antibody #9308||100 µl||W, IP, IHC-P||H, R, Mk||M||110 kDa||Rabbit|
|Rb (4H1) Mouse mAb #9309||100 µl||W, IP, IHC-P, IF-IC, F, ChIP||H, Mk, B, Pg||110 kDa||Mouse IgG2a|
|Rb Control Proteins #9303||100 µl||76 kDa|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||Goat|
|Anti-biotin, HRP-linked Antibody #7075||100 µl||Goat|
|Anti-mouse IgG, HRP-linked Antibody #7076||100 µl||Horse|
|20X LumiGLO® Reagent and 20X Peroxide #7003||5 ml each|
|Biotinylated Protein Ladder Detection Pack #7727||100 µl|
†Species predicted to react based on 100% sequence homology.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, E-P=ELISA (Peptide), IHC-P=Immunohistochemistry (Paraffin), IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry, ChIP=Chromatin IP
Reactivity Key: H=Human, R=Rat, Mk=Monkey, B=Bovine, Pg=Pig
Western blot analysis of Rb phosphorylation in human fibroblasts synchronized by serum deprivation using Phospho-Rb (Ser780) Antibody #9307 (upper) and Phospho-Rb (Ser807/811) Antibody #9308 (lower). Cells were synchronized for 24 hours then released by addition of serum and harvested at the times indicated. Cell cycle progression was verified by cyclin analysis and FACS. (Provided by John Boylan, Dupont/Merck, Delaware.)
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.
Specificity / Sensitivity
Phospho-Rb (Ser780) and Phospho-Rb (Ser807/811) antibodies detect endogenous levels of Rb only when phosphorylated at Ser780 and Ser807/811, respectively. These antibodies do not cross-react with Rb phosphorylated at other sites. 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.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues around Ser780 and Ser807/811 of human Rb. Antibodies are purified by protein A and peptide affinity chromatography. 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).
- Sherr, C.J. (1996) Science 274, 1672-7.
- Nevins, J.R. (1992) Science 258, 424-9.
- Welch, P.J. and Wang, J.Y. (1993) Cell 75, 779-90.
- Hu, Q.J. et al. (1990) EMBO J 9, 1147-55.
- Knudsen, E.S. and Wang, J.Y. (1997) Mol Cell Biol 17, 5771-83.
- Lundberg, A.S. and Weinberg, R.A. (1998) Mol Cell Biol 18, 753-61.
- Connell-Crowley, L. et al. (1997) Mol Biol Cell 8, 287-301.
- Kitagawa, M. et al. (1996) EMBO J 15, 7060-9.
- Geng, Y. et al. (2001) Proc Natl Acad Sci USA 98, 194-9.
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For Research Use Only. Not For Use In Diagnostic Procedures.
PhosphoPlus® is a trademark of Cell Signaling Technology, Inc.
LumiGLO® is a registered trademark of Kirkegaard & Perry Laboratories.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.