Description General description Purified mouse monoclonal antibody. Recognizes a broad spectrum of tyrosine-phosphorylated proteins in their native and denatured states. Recognizes a broad spectrum of tyrosine-phosphorylated proteins in their native and denatured states in crude extracts. This PhosphoDetect Anti-Phosphotyrosine Mouse mAb (PY20) is validated for use in ELISA, Immunoblotting, Frozen Sections, ICC, IP for the detection of Phosphotyrosine. Immunogen KLH conjugated to phosphotyrosine, glycine, and alanine Application ELISA (see comments) Immunoblotting (1:500-1:1000, see application references) Frozen Sections (see application references) Immunocytochemistry (see application references) Immunoprecipitation (see comments) Physical form In 10 mM sodium phosphate, pH 7.4. Reconstitution Following initial thaw, aliquot and freeze (-20°C). Packaging 100 μg in Plastic ampoule Please refer to vial label for lot-specific concentration. Warning Toxicity: Standard Handling (A) Analysis Note Positive Control Any cell lysate or tissue extract Other Notes Binding is inhibited by phosphotyrosine. Does not react with threonine or serine phosphorylated residues on proteins. Do not use milk as a blocking agent with this antibody. Milk contains many phosphotyrosines and will adsorb the antibody. This antibody has also been reported to work for ELISA and immunoprecipitation. Variables associated with assay conditions will dictate the proper working dilution. Glenney, J.R., et al. 1988. J. Immunol. Methods109, 277.