Anti-Phosphotyrosine, 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 and Immunoprecipitation., Other applications not tested. Recommended Dilution: Western Blot: 0.5-2ug/ml. Detects tyrosine-phosphorylated proteins in a modified RIPA lysate from EGF-treated human A431 carcinoma cells., Immunoprecipitation: 2-4ug. Immunoprecipitates quantitatively the phosphotyrosine-containing proteins in the lysate of a confluent culture (10cm dish) of cells expressing an activated tyrosine kinase. To preserve phosphotyrosine, add 0.2mM sodium orthovanadate to the lysis buffer., Optimal dilutions to be determined by researcher. Recommended secondary antibodies: I1904-06C: IgG, H&L (HRP) (X-Adsorbed) Pab Gt xMo, I1904-06H IgG, H&L (HRP) (X-Adsorbed) Pab Gt xMo, Positive Control (Included): A0001-55: A431 Cells, EGF-Stimulated, Storage and Stability: May be stored at 4°C. For long-term storage, aliquot and store at 4°C. Do not freeze. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made in assay buffer.