Recombinant Human Active ERK2 Protein, CF

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

Product Details

Summary
Reactivity HuSpecies Glossary
Applications Bioactivity
Format
Carrier-Free

Order Details

Recombinant Human Active ERK2 Protein, CF Summary

Details of Functionality
The activity of ERK2 is 613-829 nmol/min/mg using a myelin basic protein (MBP) substrate (see Activity Assay Protocol).
Source
E. coli-derived human ERK2 protein Accession # NM_002745
Accession #
N-terminal Sequence

Using an N terminal GST tag

Protein/Peptide Type
Recombinant Proteins
Gene
MAPK1
Purity
>70%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane

Applications/Dilutions

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

Packaging, Storage & Formulations

Storage
This product is stable at ≤ -70degrees 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, 0.25 mM DTT, 10 mM glutathione, 0.1 mM EDTA, 0.1 mM PMSF, and 25% glycerol.
Purity
>70%, 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. Note: These are suggested working dilutions. Optimal dilutions should be determined by each laboratory for each application.
  • Kinase Assay Buffer, pH 7.2 - 25 mM MOPS, 12.5 mM beta -glycerolphosphate, 25 mM MgCl2, 5 mM EGTA, 2 mM EDTA. Add 0.25 mM DTT to the Kinase Assay Buffer prior to use.
  • Kinase Dilution Buffer, pH 7.2 - Kinase Assay Buffer diluted 5-fold with distilled or deionized water.
  • 10 mM ATP Stock Solution - Prepare the ATP Stock Solution by dissolving 55 mg of ATP in 10 mL of Kinase Assay Buffer. Store 200 μL aliquots at ≤ -20° C.
  • [33P]-ATP Assay Cocktail - Prepare 250 μM [33P]-ATP Assay Cocktail in a designated radioactive work area by combining 150 μL of 10 mM ATP Stock Solution, 100 μL of [33P]-ATP (1 mCi/100 μL), and 5.75 mL of Kinase Assay Buffer. Store 1 mL aliquots at ≤ -20° C.
  • Substrate - Myelin Basic Protein (MBP) substrate diluted in distilled or deionized water to a final concentration of 1 mg/mL.
  1. Thaw the [33P]-ATP Assay Cocktail in a shielded container in a designated radioactive work area.
  2. Thaw the Active ERK2, Kinase Assay Buffer, Substrate, and Kinase Dilution Buffer on ice.
  3. In a pre-cooled microfuge tube, add the following reaction components bringing the initial reaction volume up to 20 μL.
    a. Diluted Active ERK2: 10 μL
    b. MBP Substrate (1 mg/mL; on ice): 5 μL
    c. Distilled or deionized water (on ice): 5 μL
  4. Set up the blank control as outlined in step 3, excluding the addition of the substrate. Replace the substrate with an equal volume of distilled or deionized water.
  5. Initiate the reaction with the addition of 5 μL [33P]-ATP Assay Cocktail, bringing the final volume up to 25 μL. Incubate the mixture in a water bath at 30 °C for 15 minutes.
  6. After the 15 minute incubation, terminate the reaction by spotting 20 μL of the reaction mixture onto individual pre-cut strips of phosphocellulose P81 paper.
  7. Air dry the pre-cut P81 strip and sequentially wash in a 1% phosphoric acid solution (add 10 mL of phosphoric acid to 990 mL of distilled or deionized water) with constant gentle stirring. It is recommended that the strips be washed a total of three times for approximately 10 minutes each.
  8. Count the radioactivity on the P81 paper in the presence of scintillation fluid in a scintillation counter.
  9. Determine the corrected cpm by subtracting the blank control value (see step 4) for each sample and calculate the kinase specific activity as outlined below:
    Calculation of [33P]-ATP Specific Activity (SA) (cpm/pmol)
    Specific Activity (SA) = cpm for 5 μL [33P]-ATP/pmol of ATP (in 5 μL of a 250 μM ATP stock solution; i.e. 1250 pmol)
      Calculation of Kinase Specific Activity (SA) (pmol/minutes/μg or nmol/minutes/mg)
      Corrected cpm from reaction / [(SA of 33P-ATP in cpm/pmol) x (Reaction time in minutes) x (Enzyme amount in μg or mg)] x [(Reaction volume) / (Spot Volume)]

    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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    Publications for ERK2 (1230-KS)(4)

    We have publications tested in 3 confirmed species: Human, Mouse, N/A.

    We have publications tested in 2 applications: Bioassay, EnzAct.


    Filter By Application
    Bioassay
    (3)
    EnzAct
    (1)
    All Applications
    Filter By Species
    Human
    (1)
    Mouse
    (1)
    N/A
    (1)
    All Species

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    Bioinformatics

    Gene Symbol MAPK1
    Entrez
    Uniprot