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JNK (c-Jun N-terminal kinase), also called stress activated protein kinase (SAPK), is a member of the serine/threonine MAP kinase family. MAP kinase signaling cascades are critically important in translating the signals received at the plasma membrane into changes in cellular physiology and gene expression. JNK is activated in response to a variety of stimuli, including inflammatory cytokines, growth factors and cellular stresses such as UV-light. Because of JNK’s role in basic cellular processes such as inflammation and apoptosis, an understanding of the JNK signaling cascade should speed the development of therapeutics.
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.
Antibody Specificities
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.
