| 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 # | P4078-06 |
| Applications | Suitable for use in Western Blot and Immunoprecipitation. Other applications not tested. |
| Recommended Dilution | Western Blot: 0.5-1ug/ml detects proteins containing phosphotyrosine residues in RIPA lysates from EGF-stimulated human A431 carcinoma cells. EGF-stimulatedA431 cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with anti-Phosphotyrosine (0.5ug/ml). Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system. |
| Immunoprecipitation: 4ug immunoprecipitates proteins containing phosphotyrosine residues from a human A431 RIPA lysate. |
| Optimal dilution determined by the researcher. |
| Positive Antigen Control | A0001-55 |
| EGF-stimulated A431 cell lysate. Use 20ug per lane for minigels. |
| Storage and Stability | May be stored at 4°C for short-term only. For long-term storage, store 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. |
| Clone Type | Polyclonal |
| Isotype | IgG |
| Host | Rabbit |
| Concentration | ~0.1mg/ml |
| Form | Supplied as a liquid in 0.2M Tris-glycine, pH 7.2, 0.15M sodium chloride, 1mg/ml BSA, 0.05% sodium azide, before the addition of glycerol to 40%. |
| Purity | Purified by Immunoaffinity chromatography |
| Immunogen | Rabbits were immunized with three phosphorylated immunogens: (1) phosphotyrosine covalently linked to KLH; (2) the c-Src carboxyl terminal regulatory phosphopeptide (TST EPQ-pY-QPGENL; covalently linked to KLH, and; (3) the peptide RRLIEDAE-pY-AARG; covalently linked to KLH. |
| Specificity | All of the phosphotyrosine immunoreactivity present in the antisera is immunoadsorbed. The antibody is not sequence-specific but specific for phosphotyrosine residues. |
| Species Crossreactivity | Human, mouse and rat. |
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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. |
