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c-Jun is a component of the transcription factor AP-1, which binds and activates transcription at TRE/AP-1 elements. The c-Jun oncogene encodes a nuclear protein, p39, that interacts with the AP-1 c-Fos oncogene product and forms a transacting heterodimer. c-Jun plays an important role in the regulation of gene expression and signal transduction processes and is also suggested to be actively involved in the differentiation of B-chronic lymphocytic leukemia cells and Hodgkin’s Reed-Sternberg cells as well as in the malignant transformation of melanocytes, which makes it a promising drug target.
Figure 1: Measurement of phosphorylated and total c-Jun.
NIH/3T3 cells were cultured in 96-well plates and serum-starved for 16 hours. Cells were then treated with 25 µg/ml of anisomycin for 30 minutes and fixed. Total and phospho c-Jun were each assayed in triplicate using either the phospho-c-Jun (S63), phospho-c-Jun (S73) or total c-Jun antibodies included in the FACE c-Jun Kit. Data was plotted after correction for cell number (performed through use of Crystal Violet).
Antibody Specificities
The phospho-c-Jun (S63) antibody is specific for phosphorylated c-Jun (S63) and was raised against a synthetic phospho-peptide corresponding to residues surrounding Serine 63 of human c-Jun. This antibody recognizes c-Jun when phosphorylated at this site and does not recognize the phosphorylated forms of JunD or JunB. The phospho-c-Jun (S73) antibody is specific for phosphorylated c-Jun (S73) and was raised against a synthetic phospho-peptide corresponding to residues surrounding Serine 73 of human c-Jun. This antibody recognizes c-Jun only when phosphorylated at this site and also recognizes JunD when phosphorylated at Serine 100. The total-c-Jun antibody recognizes c-Jun proteins regardless of the phosphorylation state. Each antibody is provided in a quantity sufficient for one 96-well assay.
