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The influence of lysis buffer composition on the expression and activity of protein tyrosine phosphatase
1Department of Molecular Medicine and Haematology, University of the Witwatersrand, Medical School, Johannesburg, South Africa. jce@twrinet.twr.ac.za
Abstract:
Lysis conditions are crucial in the extraction and solubilization of phosphotyrosine-containing proteins in a form that is immunoreactive, undegraded and enzymatically active. To establish optimal experimental conditions, we evaluated protein tyrosine phosphatase enzyme activity and the detection of PTP-1 B protein in human acute promyelocytic leukaemic cells, both before and after retinoic acid treatment. We found that the composition of the lysing buffer greatly influenced the efficiency of solubilization, resulting in major alterations in the activity of protein tyrosine phosphatases and on the mobility of PTP-1 B protein.