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9224
ATF-2 Fusion Protein
Experimental Controls

ATF-2 Fusion Protein #9224

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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).

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.

Supporting Data

MW (kDa) 34

Application Key:

  • W-Western
  • IP-Immunoprecipitation
  • IHC-Immunohistochemistry
  • ChIP-Chromatin Immunoprecipitation
  • IF-Immunofluorescence
  • F-Flow Cytometry
  • E-P-ELISA-Peptide

Species Cross-Reactivity Key:

  • H-Human
  • M-Mouse
  • R-Rat
  • Hm-Hamster
  • Mk-Monkey
  • Mi-Mink
  • C-Chicken
  • Dm-D. melanogaster
  • X-Xenopus
  • Z-Zebrafish
  • B-Bovine
  • Dg-Dog
  • Pg-Pig
  • Sc-S. cerevisiae
  • Ce-C. elegans
  • Hr-Horse
  • All-All Species Expected

Product Description

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.

Molecular Formula:

Molecular Weight: 34 kDa

Product Usage Information

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).

Storage:

Store at -20°C.

Quality Control

The purified protein was identified by SDS-PAGE and western blot analysis, using ATF-2 Antibody #9222, to be greater than 90% ATF-2 Fusion Protein.

Source / Purification

Cloned from a human cDNA library (5) and overexpressed in E. coli.

Background

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).

  1. Abdel-Hafiz, H.A. et al. (1992) Mol Endocrinol 6, 2079-89.
  2. Gupta, S. et al. (1995) Science 267, 389-93.
  3. van Dam, H. et al. (1995) EMBO J 14, 1798-811.
  4. Livingstone, C. et al. (1995) EMBO J 14, 1785-97.

Pathways & Proteins

Explore pathways + proteins related to this product.

For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

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