Preparation And Storage
Recommended Assay Procedures
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.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources 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.
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 (p85α). It reacts with human, monkey, and cow PI3 kinase. It does not cross-react with mouse or rat PI3 kinase. Recombinant cow p85α was used as immunogen. The epitope has been mapped to the inter-SH2 spacer region of p85α. The antibody will block lipids binding to this region.
Development References (2)
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Shoelson SE, Sivaraja M, Williams KP, Hu P, Schlessinger J, Weiss MA. Specific phosphopeptide binding regulates a conformational change in the PI 3-kinase SH2 domain associated with enzyme activation. EMBO J. 1993; 12(2):795-802. (Biology). View Reference
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Whitman M, Downes CP, Keeler M, Keller T, Cantley L. Type I phosphatidylinositol kinase makes a novel inositol phospholipid, phosphatidylinositol-3-phosphate. Nature. 1988; 332(6165):644-646. (Biology). View Reference
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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
