ERK2, GST-Tag Recombinant

Catalog #

40299

$410 *

Size: 10 µg

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Description

Recombinant full length human ERK2 (extracellular signal-regulated kinase 2). This construct contains an N-terminal GST-tag. The recombinant protein was affinity purified and was activated by MEK1 in vitro.

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Synonyms

MAPK1, P42MAPK, MAPK2, PRKM2, PRKM1, p41mapk, ERT1, p40, p41, ERK, ERK-2

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Product Data Gallery

4-20% SDS-PAGE Coomassie Staining

Specific Activity

ERK2 Activity (Testing may vary between lots.)

Product Info

Storage and Usage

Citations

Species

Human

Construct

ERK2 (GST-1-360)

Host Species/Expression System

E. coli

Purity

≥80%

Format

Aqueous buffer solution

Formulation

50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 10 mM glutathione, 0.25 mM DTT, 0.1 mM EDTA, 0.1 mM PMSF, and 25% glycerol

68 kDa

Amino Acids

1-360

Specific Activity

24 pmol/min/µg (specific activity measured using the specific activity assay described below)

Genbank #

NM_002745

UniProt #

P28482

Tag(s)

N-terminal GST-tag

Background

ERK2 is a protein serine/threonine kinase that is a member of the extracellular signal-regulated kinases (ERKs) which are activated in response to numerous growth factors and cytokines (1). Activation of ERK2 requires both tyrosine and threonine phosphorylation that is mediated by MEK. ERK2 is ubiquitously distributed in tissues with the highest expression in heart, brain and spinal cord. Activated ERK2 translocates into the nucleus where it phosphorylates various transcription factors (e.g., Elk-1, c-Myc, c-Jun, c-Fos, and C/EBP beta).

References

1. Girault, J.A., et al. Curr Opin Pharmacol. Feb,7(1):77-85 (2007)
2. Fyffe-Maricich SL., et al. J. Neurosci. Jan 19,31(3):843-50 (2011).
3. Ossum, C.G., et al. Cell Physiol Biochem. 27(2):109-20 (2011).

Storage/Stability

At least 6 months at -80°C.

Assay Conditions

Specific Activity was measured by using MBP, diluted in 100 mM MOPS, pH 6.5 to a working concentration of 0.5 mg/ml, in the ADP Glo™ Kinase Assay kit (Promega #V9101). Reaction was initiated by mixing increasing amounts of the ERK2 with 25 μM ATP in 40 mM Tris-HCl, pH 7.4, 20 mM MgCl₂, 0.1 mg/ml BSA prepared with 50 μM DTT, and substrate at a final concentration of 100 µg/ml
After a 40-minute incubation at room temperature, the reaction was terminated by addition of ADP-Glo™ Reagent, followed by a subsequent 40-minute incubation at room temperature. Kinase Detection Reagent was added, and the reaction was incubated for another 30 minutes at ambient temperature. Detection of luminescence was measured using the Luminescence Module Protocol on GloMax®-Multi Microplate Multimode Reader. The Specific Activity was calculated as follows: (Corrected activity, RLU) / [(Specific activity from ADP in RLU/pmol) * (Reaction time in min) *(Enzyme amount in μg or mg)]. Corrected RLU was calculated by subtracting the blank value from all the values. The blank was determined from a “no enzyme” sample by replacing the enzyme solution with an equal volume of Dilution Buffer IX (1x).

ERK2 Activity was measured according to Chemi-Verse™ ERK2 Kinase Assay Kit (BPS Bioscience #78899) with various amounts of ERK2.

Instructions for Use

Thaw on ice and gently mix prior to use. DO NOT VORTEX. Perform a quick spin before opening. Aliquot into small volumes and flash freeze for long term storage. Avoid multiple freeze/thaw cycles.

Applications

Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.

Shipping Temperature

-80°C

Warnings

Avoid freeze/thaw cycles

Scientific Category

Kinase/Serine-Threonine

Regulatory Status

Research Use Only

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Datasheets/Protocols

40299 Lot 230627

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