Recombinant Human Active ERK2 Protein, CF

R&D Systems | Catalog # 1230-KS

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Key Product Details

  • R&D Systems E. coli-derived Recombinant Human Active ERK2 Protein (1230-KS)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

E. coli

Accession Number

NM_002745

Applications

Bioactivity
View all ERK2 Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human ERK2 protein

Purity

>80%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.

N-terminal Sequence Analysis

Using an N terminal GST tag

SDS-PAGE

68 kDa

Activity

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.

Scientific Data Images for Recombinant Human Active ERK2 Protein, CF

Recombinant Human Active ERK2 Protein SDS-PAGE

Recombinant Human Active ERK2 Protein SDS-PAGE

The approximate molecular weight is 68 kDa and the purity is > 80%.

Formulation, Preparation, and Storage

1230-KS
Formulation 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.
Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Background: ERK2

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

Long Name

Extracellular Signal-regulated Kinase 2

Alternate Names

ERT1, MAPK1, MAPK2, p41mapk, p42mapk, PRKM1, PRKM2

Entrez Gene IDs

5594 (Human); 26413 (Mouse); 116590 (Rat)

Gene Symbol

MAPK1

UniProt

NM_002745 (Human)

Additional ERK2 Products

Product Documents for Recombinant Human Active ERK2 Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant Human Active ERK2 Protein, CF

For research use only

Citations for Recombinant Human Active ERK2 Protein, CF

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Protocols

View specific protocols for Recombinant Human Active ERK2 Protein, CF (1230-KS):

Materials
  • 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)]

    FAQs for Recombinant Human Active ERK2 Protein, CF

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    • Q: What is the activation protocol for this enzyme?

      A: This enzyme is supplied in its activated form, so no activation step is required.

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