Fast Activated Cell-based ELISA (FACE™) Kits provide a simple, sensitive method for detecting protein phosphorylation directly in the cell, without making extracts or performing electrophoresis and membrane blotting. These 96-well, high-throughput assays are available in both colorimetric and chemiluminescent formats for over 20 different targets (see list at right). For complete details, click the FACE™ Method tab below.
FACE JNK Kits provide 96 rxns each of 2 antibodies that enable you to monitor and compare the levels of both phosphorylated and total JNK (c-Jun N-terminal Kinase). The phospho-JNK antibody recognizes JNK1 only when phosphorylated at Thr183/Tyr185. The total-JNK antibody recognizes JNK1, JNK2 and JNK3 regardless of their phosphorylation states. Click the JNK Info tab below for data and more information.
Figure 1: Colorimetric measurement of phosphorylated and total JNK.
Murine macrophage 4/4 cells were cultured in 96-well plates and serum-starved for 16 hours before stimulation. Cells were stimulated at the indicated concentrations of anisomycin for 15 minutes and then fixed. Total and phospho-JNK were each assayed in triplicate using the phospho and total JNK antibodies from the FACE JNK Kit. Data was plotted after correction for cell number (performed through use of Crystal Violet). Note that the induction treatment did not affect the overall level of total JNK.
The phospho-JNK antibody is specific for phosphorylated JNK and was raised against a phospho-peptide that corresponds to the sequence surrounding Thr183/Tyr185 of human JNK. It recognizes JNK1 only when phosphorylated at Thr183/Tyr185, and JNK2 and JNK3 only when phosphorylated at equivalent sites. The total-JNK antibody recognizes JNK1, JNK2 and JNK3 regardless of the phosphorylation state.
JNK (c-Jun N-terminal kinase),