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 ATF-2 Kits provide 96 rxns each of 2 antibodies that enable you to monitor and compare the levels of both phosphorylated and total ATF-2. The phospho-ATF-2 antibody recognizes ATF-2 when dually phosphorylated at Thr69/71 as well as when singly phosphorylated at Thr71; the total-ATF-2 antibody recognizes ATF-2 regardless of the phosphorylation state. Click the ATF-2 Info tab below for data and more information.
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).
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.
ATF-2 is a member of the ATF/CREB family of leuci