Product Pathways - Motif Antibodies
Phospho-(Tyr) p85 PI3K Binding Motif Antibody #3821
| Applications | Reactivity | Source |
|---|---|---|
| W IP E-P | All | Rabbit |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
E-P=ELISA (Peptide)
Reactivity Key: All=All species expected
Specificity / Sensitivity
Phospho-(Tyr) p85 PI3K Binding Motif Antibody preferentially recognizes peptides and proteins containing phospho-tyrosine at the consensus YXXM motif. Some cross-reactivity is observed with peptides that contain phospho-tyrosine followed by methionine. The antibody does not cross-react with the corresponding nonphosphorylated sequences or with other phospho-tyrosine-containing motifs. By ELISA, the antibody recognizes a wide range of tyrosine-phosphorylated peptides with the YXXM sequence. It also recognizes proteins with phosphorylated p85 binding motifs by Western blot analysis. (Patented, U.S. No. 6,441,140 and Patents Pending.)
Source / Purification
Polyclonal antibodies are produced by immunizing rabbits with synthetic phospho-(Tyr) p85 binding motif peptides. Antibodies are sequentially purified by protein A and peptide affinity chromatography.
Western Blotting
Western blot analysis of extracts from A431 cells, untreated or sodium vanadate-treated using Phospho-(Tyr) p85 PI3K Binding Motif Antibody.
Western Blotting
Western blot analysis of extracts from untreated or EGF-treated A431 cells (detects phosphorylated EGFR), untreated or FGF-treated SK-N-MC cells (detects phosphorylated FGFR), and untreated or PDGF-treated NIH/3T3 cells(detects phosphorylated PDGFR), using Phospho-(Tyr) p85 PI3K Binding Motif Antibody.
IP
Immunoprecipitation of extracts from untreated or PDGF-treated NIH/3T3 cells, using Phospho-(Tyr) p85 PI3K Binding Motif Antibody, followed by Western immunoblotting with the same antibody (upper) or Phospho-PDGF Receptor beta (Tyr751) Antibody #3161 (lower).
ELISA-Peptide
Phospho-(Tyr) p85 PI3K Binding Motif Antibody DELFIA® Assay: Signal-to-noise ratio of phospho-versus nonphospho-peptides. (Y* denotes phosphorylated tyrosine.) (DELFIA® is a registered trademark of PerkinElmer, Inc.)
Background
Phosphoinositide 3 kinase (PI3K) is a lipid kinase that catalyzes the phosphorylation of PI(4,5)P2 at the D3 position of the inositol ring. The resulting product, PI(3,4,5)P3, can be recognized by PH domain-containing signaling molecules, including various kinases and their regulators, and activates the corresponding cell signaling pathways, eventually leading to cell growth, survival and cellular movement (1). PI3K is a heterodimer with a p85 regulatory subunit and p110 catalytic subunit. The p85 subunit has two SH2 domains that bind to phospho-tyrosine with the consensus motif (Y*XXM), where X denotes any amino acid and Y* is phosphorylated tyrosine. This motif exists in many of the receptor tyrosine kinases and their substrates, including EGFR, FGFR, IRS-1, BCAP and GAB1 (2,3). The docking of PI3K in the vicinity of the plasma membrane through p85 recognition of the receptor phosphorylated YXXM activates the PI3K pathway. Phospho-(Tyr) p85 PI3K Binding Motif Antibody specifically recognizes PI3K binding targets at their phosphorylated YXXM motif, providing a powerful new tool for PI3K binding protein discovery and characterization as well as HTS drug screening for potential regulators.
- Cantley, L.C. (2002) Science 296, 1655-1657.
- Songyang, Z. et al. (1994) Mol. Cell. Biol. 14, 2777-2785.
- Songyang, Z. et al. (1993) Cell 72, 767-778.
Application References
- Li, Y. et al. (2003) Morphine induces desensitization of insulin receptor signaling. Mol. Cell. Biol. 23 (17), 6255-6266. This article references the use of Phospho-(Tyr) p85 PI3K Binding Motif Antibody in the following applications: Western Blotting
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License/Use Restrictions: The purchase of this product does not include the right or license to practice any technology or methods described and claimed in U.S. Patent Number 7,198,896 and foreign equivalents, "Immunoaffinity Isolation of Modified Peptides from Complex Mixtures ", Rush et al., Pub. Date March 6, 2003 (and equivalents). Use of purchased product in such methods, in any field, requires a separate license from Cell Signaling Technology, Inc. A limited license to use such methods may, however, be obtained by non-profit entities via purchase of any PhosphoScan™ Kit.
