Product Pathways - Cell Cycle / Checkpoint
Rb (4H1) Mouse mAb #9309
| Applications | Reactivity | MW (kDa) | Source | Isotype |
|---|---|---|---|---|
| W IP IHC-P IF-IC F | H Mk B | 110 | Mouse | IgG2a |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IHC-P=Immunohistochemistry (Paraffin)
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key:
H=Human
Mk=Monkey
B=Bovine
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.
Specificity / Sensitivity
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
Monoclonal antibody is produced by immunizing mice with Rb-C Fusion Protein #6022, containing residues 701-928 of human Rb (KLH-coupled).
Western Blotting
Western blot analysis of extracts from COS-7 cells, untreated or hydroxyurea-treated (G1/S), using Rb (4H1) Mouse mAb.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing nuclear localization, using Rb (4H1) Mouse mAb.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Rb (4H1) Mouse mAb.
Flow Cytometry
Flow cytometric analysis of Jurkat cells, using Rb (4H1) Mouse mAb versus propidium iodide (DNA content). The box indicates Rb positive cells.
IF-IC
Confocal immunofluorescent image of SH-SY5Y cells, using RB (4H1) Mouse mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).
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
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Companion Products
- 9301 Phospho-Rb (Ser795) Antibody
- 9307 Phospho-Rb (Ser780) Antibody
- 9308 Phospho-Rb (Ser807/811) Antibody
- 2181 Phospho-Rb (Ser608) Antibody
- 6022 Rb-C Fusion Protein
- 9303 Rb Control Proteins
- 7076 Anti-mouse 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
