Comparison of Anti-Phosphotyrosine Antibodies
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 The immunochemical determination of the phosphorylation status of tyrosine is a standard technique. Which antibody to choose? Above shown you see a comparison of six different mouse antibody clones for the detection of phosphorylated tyrosine by western blotting. Phosphorylation and dephosphorylation of cellular proteins are central steps in transducing extracellular signals to the cell nucleus. Phosphorylated epitopes may serve as docking sites for the assembly of protein complexes or may alter the 3-dimensional protein structure thus modulating enzymatic activity or the ability to undergo protein-protein-interactions. Modification of proteins on tyrosine residues is mediated by protein tyrosine kinases. Tyrosine phosphorylation may alter the biological activity or mediate the assembly of protein complexes via interaction of phosphotyrosine residues with SH2 or PID domains. Anti-Phosphotyrosine clone 2C8 recognizes a broad range of tyrosine-phosphorylated proteins in crude cell extracts and may therefore be particularly well-suited for the detection/screening of tyrosine phosphorylated proteins. This clone can be used not only for western blotting, but also for immunoprecipitation and ELISA. The Fluorescein-conjugated version of Anti-Phosphotyrosine clone 2C8 can be used in flow cytometry (FC/FACS). Anti-Phosphotyrosine clone 1F9 recognizes phosphotyrosine in the context of the surrounding amino acids, tolerating charged amino acids directly neighboured to phosphotyrosine. Anti-Phosphotyrosine clone 2A5 (epitope: ... E - pY - M ...) recognizes a broad spectrum of tyrosine phosphorylated proteins in crude cell extracts. Mab 2A5 tolerates a negative charge N-terminal to the phosphotyrosine residue. Anti-Phosphotyrosine clone 3B12 recognizes phosphorylated tyrosine in immunocytochemistry (ICC). Anti-Phosphotyrosine clone 9H8 (epitope: ... R - G - pY - V - P ...) recognizes phosphotyrosine in the context of the surrounding amino acids, tolerating positively charged amino acids N-terminal to phosphotyrosine. Anti-Phosphotyrosine clone 16F4 (epitope: ... D - I - pY - A - E ...) recognizes phosphotyrosine in the context of the surrounding amino acids, tolerating hydrophobic amino acids directly neighboured to phosphotyrosine. Anti-Phosphotyrosine clone 9F1 recognizes phosphotyrosine in the context of the surrounding amino acids, tolerating hydrophobic amino acids and phosphotyrosine directly neighboured to phosphotyrosine. If you want to test several clones of antibodies for detection of phosphorylated tyrosine in your system, please choose NanoTools' Phosphotyrosine Detection Kit. Anti-Phosphotyrosine Antibodies by NanoTools |
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