Phosphorylation of specific tyrosine residues is an important post-translational modification for regulating the activity of most proteins. Stimulation of a variety of cell signaling pathways activates the receptor and non-receptor tyrosine kinases that mediate these protein modifications. Antibodies that can detect phosphotyrosine residues are excellent tools for characterizing changes in the post-translational state of a broad range of phosphotyrosine-containing proteins. Immunoprecipitation of proteins of interest, followed by detection of phosphotyrosine using anti-phosphotyrosine antibody is commonly used to correlate changes in tyrosine phosphorylation state with alterations in protein activity.
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Clone M375 was generated from tyrosine phosphorylated proteins purified from pervanadate treated A431 cells.
Western blot of HeLa cells treated with pervanadate (1 mM) for 30 min. Phosphotyrosine containing proteins were immunoprecipitated with rabbit polyclonal anti-Phosphotyrosine:Agarose (Lane 1) or with no antibody agarose beads (Lane 2), and blots were made that included the whole lysate (Lane 3). The blots were probed with mouse monoclonal anti-Phosphotyrosine (PM3751) to detect phosphotyrosine containing proteins.
Immunocytochemical labeling of phosphotyrosine in control and pervanadate-treated A431 cells. The cells were labeled with rabbit polyclonal anti-Phosphotyrosine (PP2221) and mouse monoclonal anti-Phosphotyrosine (PM3751), then the antibodies were detected using appropriate secondary antibodies conjugated to Cy3.
Mouse monoclonal antibody purified with protein A chromatography is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.
This antibody detects phosphotyrosine-containing proteins on SDS-PAGE immunoblots of EGF treated A431 cells, as well as pervanadate treated A431, HeLa, Jurkat, and endothelial cells.
*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot
mobilities of known proteins with similar MW.
This kit contains: