Anti-Phosphotyrosine, Biotin conjugated, clone 0.T.162

Protein tyrosine phosphorylation has been the subject of intense investigation for over two decades. Tyrosine phosphorylation is catalyzed by members of the tyrosine kinase superfamily. To date, approximately 90 members of this superfamily have been identified in the human genome. Levels of phosphorylation of tyrosine residues within cellular proteins are observed to increase in response to growth factors, cytokines, insulin, extracellular matrix components, and other stimuli. Tyrosine phosphorylation has several effects, including the regulation of enzyme activity, as well as the localization of enzyme activity through the generation of docking sites for proteins bearing SH2 domains. Applications: Suitable for use in Western Blot, Immunoprecipitation and Immunocytochemistry. , Other applications not tested. Recommended Dilution: Western Blot: 0.5-2ug/ml will detect tyrosine phosphorylated proteins in a RIPA lysate of EGF-stimulated A431 carcinoma cells. , Immunoprecipitation: 4ug, used in conjunction with Streptavidin-Agarose, will immunoprecipitate phosphotyrosine containing proteins from a lysate of EGF-stimulated A431 cells, to preserve phosphotyrosine, add 0.2mM sodium orthovanadate to the lysis buffer. , Immunocytochemistry: 5-10ug/ml will give a positive immunostaining of EGF-stimulated A431 cells that have been fixed with ethanol:acetic acid., Optimal dilutions to be determined by the researcher. Positive Control: Pervanadate-treated human A431 cell extracts or EGF-treated human A431 cells, Storage and Stability: Store product at 4°C if to be used immediately within two weeks. Do not freeze. For long-term storage, aliquot to avoid repeated freezing and thawing and store at 4°C. Aliquots are stable at 4°C for 12 months after receipt. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.