Activation of
protein kinase, such as JNK and Akt, plays an important role
in apoptosis and other physiological and pathological
processes. BioVision provides a novel nonradioactive
system for convenient measurement of JNK and Akt activities.
The JNK Activity Screening Kit is used to screen JNK
activity relatively quickly for large numbers of samples.
The assay utilizes a N-terminal c-Jun fusion protein bead to
selectively “pull down” JNK from cell lysate. c-Jun
phosphorylation is then measured using a phospho-c-Jun
specific antibody. The KinaseSTARTM JNK
or Akt Activity Assay Kit utilizes a JNK- (or Akt-)
specific antibody to immunoprecipitate JNK (or Akt) from
cell lysate. JNK- (or Akt-) specific activity is then
analyzed by Western blot analysis of the phosphorylation
levels of c-Jun (or GSK-3a) using a phospho-c-Jun (or
phospho-GSK-3a) specific antibody. The kit specifically
detects JNK (or Akt) activity, other kinase activities would
not be detected.
Analyses of JNK Specific Activity in
Jurkat Extracts. Jurkat cells were untreated (Lane 3),
or treated with anisomycin (10 ug/ml, Lane 1) or NaCl (0.5
M, Lane 2) for 30 min. Cell lysates were immunoprecipitated
using a JNK-specific polyclonal antibody. JNK specific
activity was then analyzed by Western blot analysis of the
phosphorylation levels of c-Jun (1-79), using a phospho-c-Jun
specific antibody included in the kit.
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