Recombinant Human Active ERK2 Protein, CF

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Recombinant Human Active ERK2 Protein SDS-PAGE
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Product Details
Citations (4)

Recombinant Human Active ERK2 Protein, CF Summary

Product Specifications

>80%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.
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.
E. coli-derived human ERK2 protein
Accession #
N-terminal Sequence
Using an N terminal GST tag
68 kDa

Product Datasheets

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Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.


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.

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)]

    Scientific Data

    SDS-PAGE Recombinant Human Active ERK2 Protein SDS-PAGE View Larger

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

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    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 ).

    1. Boulton, T.G. et al. (1991) Biochemistry 30(1):278.
    Long Name
    Extracellular Signal-regulated Kinase 2
    Entrez Gene IDs
    5594 (Human); 26413 (Mouse); 116590 (Rat)
    Alternate Names
    EC 2.7.11; EC; 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

    Citations for Recombinant Human Active ERK2 Protein, CF

    R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

    4 Citations: Showing 1 - 4
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    1. Deregulated PP1? phosphatase activity towards MAPK activation is antagonized by a tumor suppressive failsafe mechanism
      Authors: M Chen, L Wan, J Zhang, J Zhang, L Mendez, JG Clohessy, K Berry, J Victor, Q Yin, Y Zhu, W Wei, PP Pandolfi
      Nat Commun, 2018;9(1):159.
      Applications: Bioassay
    2. IL-1? induced and p38(MAPK)-dependent activation of the mitogen-activated protein kinase (MAPK)-activated protein kinase (MK) 2 in hepatocytes: signal transduction with robust and concentration-independent signal amplification
      Authors: A Kulawik, R Engesser, C Ehlting, A Raue, U Albrecht, B Hahn, WD Lehmann, M Gaestel, U Klingmülle, D Häussinger, J Timmer, JG Bode
      J. Biol. Chem, 2017;0(0):.
      Species: Human
      Sample Types: Recombinant Protein
      Applications: Enzyme Assay
    3. Effective combinatorial immunotherapy for castration-resistant prostate cancer
      Authors: X Lu, JW Horner, E Paul, X Shang, P Troncoso, P Deng, S Jiang, Q Chang, DJ Spring, P Sharma, JA Zebala, DY Maeda, YA Wang, RA DePinho
      Nature, 2017;543(7647):728-732.
      Species: Mouse
      Sample Types: Whole Cells
      Applications: Bioassay
    4. PPARgamma recruitment to active ERK during memory consolidation is required for Alzheimer's disease-related cognitive enhancement.
      Authors: Jahrling J, Hernandez C, Denner L, Dineley K
      J Neurosci, 2014;34(11):4054-63.
      Species: N/A
      Sample Types: Beads
      Applications: Bioassay


    1. What is the activation protocol for this enzyme?

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

    View all Proteins and Enzyme FAQs

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