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. Other applications not tested. Recommended Dilution: Western Blot: 0.5-2ug/ml detected tyrosine phosphorylated proteins in a RIPA lysate of EGF-stimulated A431 carcinoma cells., Optimal dilutions to be determined by the researcher. Included Positive Antigen Control: EGF-stimulated A431 cell lysate is provided as a free positive antigen control for Western immunoblotting. Aliquot as desired, refreeze immediately, and store at -20°C. The lysate is stable for 6 months at -20°C. Before use, add 2.5ul of 2-mercaptoethanol/100ul of lysate and boil for 5 minutes to reduce the preparation. Load 20ug of reduced lysate per lane for immunoblot analysis. Storage and Stability: Store product at 4°C if to be used immediately within two weeks. For long-term storage, aliquot to avoid repeated freezing and thawing and store at -20°C. Aliquots are stable at -20°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.