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9184
Elk-1 Fusion Protein
Experimental Controls

Elk-1 Fusion Protein #9184

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Figure 2. Elk-1 fusion protein was used as substrate to measure Erk1 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: Elk-1 fusion protein 400 ng/μL, and variable amounts of Erk1 kinases.

Supporting Data

MW (kDa) 41, 45

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

Elk-1 Fusion Protein serves as a useful substrate for p42/44 MAP kinases (1,6). It is expressed as a recombinant protein fusion containing Elk-1 residues 307–428.

Molecular Formula:

Apparent Molecular Weight: 41 kDa

Bioactivity:

Elk 1 Fusion Protein at a concentration of 2 µg/20 µ| reaction was phosphorylated using p42 MAP kinase (Erk1) in an in vitro kinase assay with 1X Kinase Buffer #9802 and 200 µM ATP #9804. After a 30-minute assay at 30°, phosphorylation was detected by Western blot with Phospho-Elk-1 (Ser383) Antibody #9181.

Product Usage Information

Elk 1 Fusion Protein, at a concentration of 2 µg/20 µl reaction, can be phosphorylated using 50 units of p42 MAP kinase (Erk2) 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-Elk-1 (Ser383) Antibody #9181.

Storage:

Store at -20°C.

Quality Control

The purified fusion protein was identified by SDS-PAGE and Western blot analysis, using Elk-1 Antibody #9182, to be greater than 95% pure Elk-1 .

Source / Purification

Cloned from a human cDNA library (7) and overexpressed in E. coli.d in E. coli.

Background

The transcription factor Elk-1 is a component of the ternary complex that binds the serum response element (SRE) and mediates gene activity in response to serum and growth factors (1-3). Elk-1 is phosphorylated by MAP kinase pathways at a cluster of S/T motifs at its carboxy terminus; phosphorylation at these sites, particularly Ser383, is critical for transcriptional activation by Elk-1. Elk-1 appears to be a direct target of activated MAP kinase: (a) biochemical studies indicate that Elk-1 is a good substrate for MAP kinase; (b) the kinetics of Elk-1 phosphorylation and activation correlate with MAP kinase activity; (c) interfering mutants of MAP kinase block Elk-1 activation in vivo. Other studies have shown that Elk-1 (Ser383) is also a target of the stress-activated kinase SAPK/JNK (4,5).

  1. Marais, R. et al. (1993) Cell 73, 381-93.
  2. Kortenjann, M. et al. (1994) Mol Cell Biol 14, 4815-24.
  3. Hill, C.S. and Treisman, R. (1995) Cell 80, 199-211.
  4. Cavigelli, M. et al. (1995) EMBO J. 14, 5957-5964.
  5. Whitmarsh, A.J. et al. (1995) Science 269, 403-407.

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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