METHOD FOR MEASURING JNK/SAPK ACTIVITY BY PHOSPHORYLATION OF TRANSCRIPTION FACTORS

摘要

A method for determining the activity of jnk / sapks, wherein said method comprising the steps of:(a) preparation of a cell lysate or woven fabric lysate;(b) purification of endogenous jnk / sapks;(c) carrying out a kinase - reaction with the obtained after step (b) jnk / sapks in the presence of atp and one or both n - terminal phosphorylation sites containing gst - transcription factor - protein, wherein the concentration range of gst - transcription factor - protein between 0,2 and 5 μg per 50 μl kinase - reaction and of cell lysate of between 50 and 1000 μg per 50 μl kinase - reaction is;(d) detection of the amount of phosphorylated gst - transcription factor - protein by the addition of a specific antibody - p - gst - transcription factor - antibody in excess in relation to the mixture of step (c) and determining the activity of the jnk / sapks on the basis of the amount of bound antibodies - p - gst - transcription factor - antibody.