Product Pathways - Cell Cycle / Checkpoint
Phospho-Rb (Ser608) Antibody #2181
| Applications | Reactivity | MW (kDa) | Source |
|---|---|---|---|
| W IP IHC-P IF-IC F | H M R Mk (Dg) | 110 | Rabbit |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IHC-P=Immunohistochemistry (Paraffin)
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key:
H=Human
M=Mouse
R=Rat
Mk=Monkey
Dg=Dog
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.
Specificity / Sensitivity
Phospho-Rb (Ser608) Antibody detects endogenous levels of Rb only when phosphorylated at serine 608.
Source / Purification
Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Ser608 of human Rb. Antibodies are purified by protein A and peptide affinity chromatography.
Western Blotting
Western blot analysis of extracts from HeLa cells treated with lambda Phosphatase NEB #P0753 (10,000 units/ml, 1 h) or calyculin A #9902 (100 nM, 10 min) using Phospho-Rb (Ser608) Antibody (upper) or Rb (4H1) Mouse mAb #9309 (lower).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, showing nuclear localization, using Phospho-Rb (Ser608) Antibody.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human Non-Hodgkin's lymphoma, untreated (left) or lambda phosphatase-treated (right), using Phospho-Rb (Ser608) Antibody.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded U2-OS (top) or SAOS-2 (bottom) cells, untreated (left) or serum-treated (right), using Phospho-Rb (Ser608) Antibody. Note the lack of induced nuclear staining in Rb-deficient SAOS-2 cells.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Phospho-Rb (Ser608) Antibody in the presence of control peptide (left) or antigen specific peptide (right).
Flow Cytometry
Flow cytometric analysis of untreated Jurkat cells, using Phospho-Rb (Ser608) Antibody versus propidium iodide (DNA content). The boxed population indicates p-Rb (Ser608)-positive cells.
IF-IC
Confocal immunofluorescent analysis of HeLa cells labeled with Phospho-Rb (Ser608) Antibody (green) showing localization in proliferating cells. Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).
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).
- Sherr, C.J. (1996) Science 274, 1672-1677.
- Nevins, J.R. et al. (1992) Science 258, 424-429.
- Welch, P.J. and Wang, J.Y. (1993) Cell 75, 779-790.
- Hu, Q.J. et al. (1990) EMBO J. 9, 1147-1155.
- Knudsen, E.S. and Wang, J.Y. (1997) Mol. Cell. Biol. 17, 5771-5783.
- Lundberg, A.S. and Weinberg, R.A. (1998) Mol. Cell. Biol. 18, 753-761.
- Connell-Crowley, L. et al. (1997) Mol. Cell. Biol. 8, 287-301.
- Kitagawa, M. et al. (1996) EMBO J. 15, 7060-7069.
- Geng, Y. et al. (2001) Proc. Natl. Acad. Sci. USA 98, 194-199.
Application References
- von Willebrand, M. et al. (2003) The tyrphostin AG1024 accelerates the degradation of phosphorylated forms of retinoblastoma protein (pRb) and restores pRb tumor suppressive function in melanoma cells. Cancer Res 63, 1420-1429. This article references the use of Phospho-Rb (Ser608) Antibody in the following applications: Western Blotting
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Companion Products
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- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
