MAPKAPK2 (MAPKAP kinase 2) is a Ser/Thr protein kinase which is regulated via direct phosphorylation by p38 MAP kinase (1). In conjunction with p38 MAP kinase, MAPKAPK2 is known to be involved in many cellular processes including stress and inflammatory responses, nuclear export, gene expression regulation, and cell proliferation. Heat shock protein 27 (HSP27) has been shown to be one of the substrates of MAPKAPK2 in vivo.
Recombinant Human Active MAPKAPK2 (46-end) Protein, CF
R&D Systems | Catalog # 3705-KS
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Key Product Details
- R&D Systems Sf 9 (baculovirus)-derived Recombinant Human Active MAPKAPK2 (46-end) Protein (3705-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
Sf 9 (baculovirus)
Accession Number
Applications
Bioactivity
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Product Specifications
Source
Spodoptera frugiperda, Sf 9 (baculovirus)-derived human MAPKAPK2 protein
Purity
>90%, 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 MAPKAPK2 is typically 233-315 nmol/min/mg using a synthetic peptide substrate (RRLNRQLSVA-amide) (see Activity Assay Protocol).
Scientific Data Images for Recombinant Human Active MAPKAPK2 (46-end) Protein, CF
Recombinant Human Active MAPKAPK2 (46-end) Protein SDS-PAGE.
The approximate molecular weight is 68 kDa and the purity is > 90%.Formulation, Preparation, and Storage
3705-KS
| Formulation | Supplied in 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 10 mM Glutathione, 0.1 mM EDTA, 0.25 mM DTT, 0.1 mM PMSF, 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: MAPKAPK2
References
- Stokoe, D. et al. (1993) Biochem. J. 296:843.
Long Name
Mitogen-Activated Protein Kinase-Activated Protein Kinase 2
Alternate Names
MK2, Rps6kc1
Gene Symbol
MAPKAPK2
UniProt
Additional MAPKAPK2 Products
Product Documents for Recombinant Human Active MAPKAPK2 (46-end) Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human Active MAPKAPK2 (46-end) Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human Active MAPKAPK2 (46-end) Protein, CF
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Protocols
View specific protocols for Recombinant Human Active MAPKAPK2 (46-end) Protein, CF (3705-KS):
Materials
- Active Kinase - Active MAPKAPK2 (0.1 μg/μL) diluted with Kinase Dilution Buffer III and assayed as outlined in sample activity plot. Note: These are suggested working dilutions and it is recommended that the researcher perform a serial dilution of Active MAPKAPK2 for optimal results.
- Kinase Assay Buffer I - 25 mM MOPS, pH 7.2, 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 III - Kinase Assay Buffer I diluted at a 1:4 ratio (5X dilution) with 50 ng/μL BSA solution.
- 10 mM ATP Stock Solution - Prepare the ATP Stock Solution by dissolving 55 mg of ATP in 10 mL of Kinase Assay Buffer I. 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 adding the following components: 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 I. Store 1 mL aliquots at -20 °C.
- Substrate - HSP27tide synthetic peptide substrate (RRLNRQLSVA-amide) diluted in distilled water to a final concentration of 1 mg/mL.
- Thaw the [33P]-ATP Assay Cocktail in a shielded container in a designated radioactive work area.
- Thaw the Active MAPKAPK2, Kinase Assay Buffer I, Substrate, and Kinase Dilution Buffer on ice.
- In a pre-cooled microfuge tube, add the following reaction components bringing the initial reaction volume up to 20 μL:
a. Diluted Active MAPKAPK2: 10 μL
b. Substrate (1 mg/mL Stock Solution): 5 μL
c. Distilled water (2-8 °C): 5 μL - 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 water.
- Initiate the reaction by 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.
- After the 15 minute incubation period, terminate the reaction by spotting 20 μL of the reaction mixture onto individual pre-cut strips of phosphocellulose P81 paper.
- Air dry the pre-cut P81 strip and sequentially wash in a 1% phosphoric acid solution (dilute 10 mL of phosphoric acid and make a 1 liter solution with distilled water) with constant gentle stirring. It is recommended that the strips be washed a total of 3 intervals for approximately 10 minutes each.
- Count the radioactivity on the P81 paper in the presence of scintillation fluid in a scintillation counter.
- Determine the corrected cpm by removing 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)]
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Associated Pathways
