Method for measuring tyrosine kinase phosphorylation

Details Method for measuring tyrosine kinase phosphorylation Method for measuring tyrosine kinase phosphorylation

2011-07-19

2011-07-19

Gitomer, Ralph (Department: 1657)

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving transferase

C435S007940

Reexamination Certificate

active

07981633

ABSTRACT:
The present invention relates to methods for measuring the autophosphorylation of one or more tyrosine kinases and use of such methods in profiling kinase inhibitors and activators. As a representative example, the method comprises inducing kinase autophosphorylation activity in cells in presence and in absence of a kinase inhibitor, lysing the cells, capturing the tyrosine kinase in the cell lysate by adding a plurality of tyrosine kinase specific binding proteins which are associated with unique dyes, adding a phosphotyrosine specific antibody tagged with a marker which is distinguishable from the unique dyes, and identifying the autophosphorylated tyrosine kinase by detecting the unique dye and the marker. Alternately, the tyrosine kinases themselves could be coupled to the unique dyes. The present invention also relates to kits and compositions for carrying out the above-described methods.

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