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ATF-2 is a member of the ATF/CREB family of leucine zipper DNA binding proteins that regulate transcription. This family of transcription factors play an important role in cell growth and differentiation, as well as in stress and immune responses. ATF-2 is a nuclear protein that binds to DNA as a dimer, and can form both homo- and heterodimers with members of the ATF/CREB and Jun/Fos families. The ATF-2/c-Jun heterodimer acts as a stronger activator than the ATF-2 homodimer. ATF-2 plays a role in c-Jun-dependent cell cycle progression, cell survival and apoptosis, making efficient study methods in high demand.
Figure 1: Measurement of phosphorylated and total ATF-2.
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 ATF-2 were each assayed in triplicate using the phospho and total ATF-2 antibodies included in the FACE ATF-2 Kit. Data was plotted after correction for cell number (performed through use of Crystal Violet).
Antibody Specificities
The phospho-ATF-2 antibody is specific for phosphorylated ATF-2 and was raised against a synthetic phospho-peptide corresponding to residues surrounding Thr71 of human ATF-2. This antibody recognizes ATF-2 when phosphorylated at this site and does not cross-react with c-Jun, CREB or other transcription factors. This antibody recognizes both ATF-2 when dually phosphorylated at Thr69/71 as well as ATF-2 when singly phosphorylated at Thr71. The total-ATF-2 antibody recognizes ATF-2 proteins regardless of the phosphorylation state.
