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- W1006854105 abstract "This chapter describes methods for preparing the PY-protein and PYA-protein conjugates, immunizing mice, and screening hybridoma culture fluid for specific antibodies. It compares and contrasts the affinities and fine specificities of these antibodies with those of the earlier anti-ABP monoclonal antibody (2G8) and provides detailed methods for using the most avid and specific monoclonal antibody (1G2) for purifying PY-containing proteins from cells. The first polyclonal and monoclonal antibodies reactive with phosphotyrosine (PY)-containing proteins were generated by immunizing animals with p-aminobenzylphosphonic acid (ABP) diazotized to carrier protein. This analog of phosphotyrosine was chosen over phosphotyrosine as the haptenic group because the phosphonate group, in contrast to the phosphate group of PY, was resistant to enzymatic hydrolysis, and thus might be a more persistent and efficient immunogen. Polyclonal and monoclonal antibodies to ABP, however frequently cross-reacted with a number of other phosphate-containing compounds. One ABP-induced monoclonal antibody, 2G8, had been extremely useful for characterizing and purifying a variety of phosphotyrosyl proteins." @default.
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- W1006854105 date "1991-01-01" @default.
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- W1006854105 title "[8] Generation of monoclonal antibodies against phosphotyrosine and their use for affinity purification of phosphotyrosine-containing proteins" @default.
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- W1006854105 doi "https://doi.org/10.1016/0076-6879(91)01010-y" @default.
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