Description
The retinoblastoma gene encodes a nuclear phosphoprotein (Rb or p110[ Rb]which is expressed in most normal cells of vertebrates and acts as a tumor suppressor gene product. An underphosphorylated form of Rb is mainly found in resting or fully differentiated cells, whereas the hyperphosphorylated form is present in proliferating cells. Only the underphosphorylated form of Rb binds specifically to viral oncogenes such as SV40 large T, adenoviral EIA and HPV-E7. This interaction may partially contribute to the transforming activity of these viral oncoproteins. Rb also interacts with several cyclins including A, D, and E as well as the transcriptional activator E2F. The importance of these interactions for the biological function of Rb is still being elucidated. The G99-2005 antibody recognizes an epitope in the first 240 amino acids of human Rb. A truncated recombinant human TrpE-Rb fusion protein was used as immunogen.
Format
- FormatPurified
Suggested Companion Products
HRP Goat Anti-Mouse Ig RUO 1mLCat No: 554002
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Preparation and Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.Store undiluted at 4°C.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
Rb migrates as multiple closely-spaced bands between ~110-116 kDa when sized on denaturing polyacrylamide gels (i.e. by SDS-PAGE). The different bands represent different Rb phosphorylation states, the higher molecular weight bands are more highly phosphorylated than the lower molecular weight bands. The level of phosphorylation is cell cycle dependent, and may also be cell type dependent (not all forms are seen in all cell types that express Rb). Gel conditions influence the actual number of bands observed. In cases where optimal band separation is desired, use a 4 to 20% gradient long ( ≥12 inches) gel. Applications include immunoprecipitation (1-2 µg/1x10^6 cells) and western blot analysis (2 µg/ml). The antibody is routinely tested by using MOLT-4 human leukemia cells (ATCC CRL-1582).
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