Product Pathways - MAPK Signaling
ATF-2 Fusion Protein #9224
|9224L||.5 mg||---||In Stock||---|
|9224S||.1 mg||---||In Stock||---|
|9224||carrier free and custom formulation / quantity||email request|
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Activating Transcription Factor 2 (ATF-2) Fusion Protein serves as a useful substrate for SAPK/JNK and p38 MAP kinases. It is expressed as a recombinant protein fusion containing ATF-2 residues 19-96. It contains the amino-terminal activation domain of ATF-2, which is regulated by phosphorylation of Thr69 and Thr71.
Figure 1. Western blot analysis of ATF-2 fusion protein phosphorylated by different isoforms of p38 kinase, using ATF-2 (20F1) Rabbit mAb #9226 (upper panel) and Phospho-ATF-2 (Thr71) (11G2) Rabbit mAb #5112 (lower panel).
Kinase Assay - Radiometric
Figure 2. ATF-2 fusion protein was used as substrate to measure p38 kinases activity in a radiometric assay using the following reaction conditions: 25 mM Tris-HCl (pH7.5), 10 mM MgCl2, 5 mM b-glycerophosphate, 0.1 mM Na3VO4, 2 mM DTT, 50 μM ATP, Substrate: ATF-2 fusion protein 400 ng/μL, and p38 kinases: 100 ng/25 μL.
Directions For Use
ATF-2 Fusion Protein, at a concentration of 2 µg/20 µl reaction, can be phosphorylated by an upstream kinase in an in vitro kinase assay with 1X Kinase Buffer (#9802) and 200 µM ATP (#9804). After a 30-minute assay at 30ºC, phosphorylation can be detected by Western blot with Phospho-ATF-2 (Thr71) Antibody (#9221).
The transcription factor ATF-2 (also called CRE-BP1) binds to both AP-1 and CRE DNA response elements and is a member of the ATF/CREB family of leucine zipper proteins (1). ATF-2 interacts with a variety of viral oncoproteins and cellular tumor suppressors and is a target of the SAPK/JNK and p38 MAP kinase signaling pathways (2-4). Various forms of cellular stress, including genotoxic agents, inflammatory cytokines, and UV irradiation, stimulate the transcriptional activity of ATF-2. Cellular stress activates ATF-2 by phosphorylation of Thr69 and Thr71 (2-4). Both SAPK and p38 MAPK have been shown to phosphorylate ATF-2 at these sites in vitro and in cells transfected with ATF-2. Mutations of these sites result in the loss of stress-induced transcription by ATF-2 (2-4). In addition, mutations at these sites reduce the ability of E1A and Rb to stimulate gene expression via ATF-2 (2).
- Heuertz, R. M. et al. (1999) . Journal of Biological Chemistry 274, 17968-17974. Applications: kinase assay substrate.
- Page, K. et al. (1999) . American Journal of Respiratory Cell and Molecular Biology 20, 1294-1302. Applications: kinase assay substrate.
- Rao, G.N. (2000) . Journal of Biological Chemistry 275, 16993-16999. Applications: kinase assay substrate.
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