| 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. |
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| Catalog # | P4075-32B |
| 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.1,2,3. |
| Immunoprecipitation: 2-4ug. Immunoprecipitate 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 the researcher. |
| Positive Control (Included) | A0001-55: A431 Cells, EGF-Stimulated |
| Hybridoma | (FOX-NY [NS-1 derivative] myeloma x spleen cells) |
| Storage and Stability | May be stored at 4°C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%). Freeze at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. |
| CAS Number | n/a |
| Clone Type | Monoclonal |
| Isotype | IgG2b,kappa |
| Clone No | 0.T.162 |
| Host | Mouse |
| Concentration | ~1mg/ml |
| Form | Supplied as a liquid in 0.1M Tris-glycine, pH 7.4, 0.15M sodium chloride, 0.05% sodium azide. |
| Purity | Purified by Protein G affinity chromatography. |
| Immunogen | Phosphotyramine (KLH coupled) |
| Specificity | Recognizes phosphotyrosine and protein-phosphotyrosine. Does not react with phosphoserine or phosphothreonine. |
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| Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
