Western Blotting (WB), ELISA, Immunoprecipitation (IP)
Marque
BD Pharmingen™
Réactivité croisée
Humain, Singe
Attributs du produit
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results. 2. Please refer to us for technical protocols. 3. 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.
Purification
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
Clone U15 was originally characterized by ELISA, immunoprecipitation, and western blot analysis. Applications include immunoprecipitation (1-2 µg/1x10^6 cells) and western blot analysis (1-2 µg/ml). In immunoprecipitation, U15 can bring down an active kinase. Jurkat human T cells (ATCC TIB-152) are suggested as a positive control.
This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Stock
4 °C
Stockage commentaire
Store undiluted at 4°C.
Shoelson, Sivaraja, Williams, Hu, Schlessinger, Weiss: "Specific phosphopeptide binding regulates a conformational change in the PI 3-kinase SH2 domain associated with enzyme activation." dans: The EMBO journal, Vol. 12, Issue 2, pp. 795-802, (1993) (PubMed).
Whitman, Downes, Keeler, Keller, Cantley: "Type I phosphatidylinositol kinase makes a novel inositol phospholipid, phosphatidylinositol-3-phosphate." dans: Nature, Vol. 332, Issue 6165, pp. 644-6, (1988) (PubMed).
Phosphatidylinositol 3-kinase (PI3-kinase) is a universal enzyme associated with receptor signaling pathways. It generates second messenger phospholipids by phosphorylating the D-3 position of inositol phospholipids including phosphatidylinositol (PI), PI-4-phosphate, and PI-4, 5-biphosphate. The enzyme exists as a heterodimer composed of regulatory 85 kDa (p85) and catalytic 110 kDa (p110) subunits. The p85 subunit contains two SH2 (src-homology 2) domains and an SH3 domain. The catalytic activity of the p110 subunit is stimulated when the p85 regulatory subunit binds, through its SH2 domains, to activated receptor and non-receptor tyrosine kinases. Two p85 isoforms have been described, p85a and p85ß. Both isoforms bind to activated receptors and each may be responsible for mediating a subset of PI3-kinase interactions. Clone U15 recognizes the p85 regulatory subunit of PI3 kinase (p85alpha). It reacts with human, monkey, and cow PI3 kinase. It does not cross-react with mouse or rat PI3 kinase. Recombinant cow p85alpha was used as immunogen. The epitope has been mapped to the inter-SH2 spacer region of p85alpha. The antibody will block lipids binding to this region.