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Recombinant Human Active ERK2 Protein, CF

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The approximate molecular weight is 68 kDa and the average purity is 95%.
The approximate molecular weight is 68 kDa and the average purity is 95%.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications Bioactivity
Format
Carrier-Free

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Recombinant Human Active ERK2 Protein, CF Summary

Details of Functionality
The specific activity of ERK2 was determined to be 24 nmol/min/mg using a myelin basic protein (MBP) substrate and was equivalent to 639 nmol/min/mg as per radiometric assay.
Source
E. coli-derived human ERK2 protein
Accession #
N-terminal Sequence
Using an N terminal GST tag
Protein/Peptide Type
Recombinant Proteins
Gene
MAPK1
Purity
>80%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.

Applications/Dilutions

Dilutions
  • Bioactivity
SDS-PAGE
68 kDa
Publications
Read Publications using
1230-KS in the following applications:

Packaging, Storage & Formulations

Storage
This product is stable at ≤ ‑70°C for up to 1 year from the date of receipt. For optimal storage, aliquot into smaller quantities after centrifugation and store at recommended temperature. Avoid repeated freeze-thaw cycles.
Buffer
Supplied in 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.
Purity
>80%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.
Assay Procedure
  • Active Kinase - Active ERK2 (0.1 μg/μL) diluted with Kinase Dilution Buffer IX (1X) and assayed as outlined in sample activity plot. Note: These are suggested working dilutions. Optimal dilutions should be determined by each laboratory for each application.
  • Kinase Assay Buffer III (5X) - 200 mM Tris-HCl, pH 7.4, 100 mM MgCl2, and 0.5 mg/mL BSA. Add fresh DTT prior to use to a final concentration of 250 μM.
  • Kinase Dilution Buffer IX (1X) - Kinase Assay Buffer III diluted at a 1:4 ratio (5X dilution) with cold water. Add fresh DTT to the aliquot prior to use to a final concentration of 50 μM.
  • ADP-Glo™ Kinase Assay Kit - 10 mM ATP Solution, 10 mM ADP Solution, ADP-Glo™ Reagent, Kinase Detection Reagent
  • Substrate - Myelin Basic Protein (MBP) diluted in 100 mM MOPS, pH 6.5, to a final concentration of 0.5 mg/mL.
  1. Thaw the Active ERK2, Kinase Assay Buffer III (5X), and Substrate on ice. Prepare a 15 μL enzyme dilution at the desired concentration, with Kinase Dilution Buffer IX (1X), in a pre-chilled 96-well plate.
  2. Prepare a substrate/ATP mixture as follows (25 μM example):
    a. 10 mM ATP Solution: 1 μL
    b. Kinase Assay Buffer III (5X): 79 μL
    c. Substrate at 0.5 mg/mL: 80 μL
  3. Transfer the following reaction components prepared in Step 2 to a 384-well opaque plate bringing the reaction volume up to 5 μL:
    a. 3 μL of diluted Active ERK2
    b. 2 μL of Substrate/ATP mix as prepared in the table above. This initiates the reaction.
  4. Set up the blank control as outlined in step 2, excluding the addition of the kinase. Replace the kinase with an equal volume of Kinase Dilution Buffer IX (1X).
  5. Incubate at ambient temperature for 40 minutes.
  6. After the 40 minute incubation period, terminate the reaction and deplete the remaining ATP by adding 5 μL of ADP-Glo™ Reagent. Spin down and shake the 384-well plate. Then incubate the reaction mixture for another 40 minutes at ambient temperature.
  7. Add 10 μL of the Kinase Detection Reagent to the 384-well plate and incubate the reaction mixture for another 30 minutes at ambient temperature.
  8. Read the 384-well reaction plate using the Luminescence Module Protocol on a GloMax®-Multi Microplate Multimode Reader.
  9. Determine the corrected activity (RLU) by removing the blank control value (see step 4) for each sample and calculate the kinase specific activity as outlined below.
    Calculation of Specific Activity of ADP (RLU/pmol)
    From ADP standard curve, determin RLU/pmol of ADP
      Kinase Specific Activity (SA) (pmol/min/μg or nmol/min/mg)
      Corrected RLU from reaction / [(SA of ADP in RLU/pmol) x (Reaction time in minutes) x (Enzyme amount in μg or mg)]

    Notes

    This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

    Alternate Names for Recombinant Human Active ERK2 Protein, CF

    • EC 2.7.11
    • EC 2.7.11.24
    • ERK
    • ERK2
    • ERK-2
    • ERK2MAP kinase isoform p42
    • ERT1
    • Extracellular signal-regulated kinase 2
    • MAP kinase 1
    • MAP kinase 2
    • MAPK 1
    • MAPK1
    • MAPK2
    • mitogen-activated protein kinase 1
    • Mitogen-activated protein kinase 2
    • p38
    • p40
    • p41
    • p41mapk
    • p42mapk
    • p42-MAPK
    • PRKM1
    • PRKM1MAPK 2
    • PRKM2
    • protein tyrosine kinase ERK2

    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 ).

    1. Boulton, T.G. et al. (1991) Biochemistry 30(1):278.

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    Recombinant Human
    Active ERK2 Protein, ...
    1230-KS
    Recombinant Human Active ERK2 Protein, CF
    Species: Hu
    Applications: Bioactivity

    Publications for ERK2 (1230-KS)(4)

    We have publications tested in 2 confirmed species: Human, Mouse.

    We have publications tested in 2 applications: Bioassay, Enzyme Assay.


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    Bioinformatics

    Gene Symbol MAPK1
    Uniprot