MAPKAPK2, His-tag Recombinant

Catalog #

40116

$410 *

Size: 10 µg

Qty

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Description

Recombinant human full length MAPKAPK2 (MAP kinase-activated protein kinase 2). This construct contains an N-terminal His-tag. The recombinant protein was affinity purified and is active.

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Synonyms

MK2

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Product Data Gallery

4-20% SDS-PAGE Coomassie Staining

Specific Activity

Product Info

Storage and Usage

Citations

Species

Human

Construct

MAPKAPK2 (His-46-end)

Host Species/Expression System

Sf9 insect cells

Purity

≥75%

Format

Aqueous buffer solution

Formulation

50 mM Sodium Phosphate, pH 7.5, 300 mM NaCl, 150 mM Imidazole, 0.25 mM DTT, 0.1 mM PMSF, 25% glycerol

41 kDa

Amino Acids

46–end

Specific Activity

442 pmol/min/µg

Genbank #

NM_032960

UniProt #

P49137

Tag(s)

N-terminal His-tag

Background

MAPKAPK2 (MAPKAP kinase2) 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 HSP27 has been shown to be one of the substrates of MAPKAPK2, in vivo.

References

1. Stokoe, D. et al . Biochem. J. 296: 843-849, 1993.
2. Hagood JS, Olman MA. Am. J. Respir. Cell Mol. Biol. 37(5):503-6 (2007).
3. Kotlyarov A, Gaestel M. Biochem. Soc. Trans. 30(Pt 6):959-63 (2002).

Storage/Stability

At least 6 months at -80°C.

Assay Conditions

MAPKAPK2 activity was measured by using the HSP27tide synthetic peptide (RRLNRQLSVA-amide) at a working concentration of 1 mg/ml, in the ADP Glo™ Kinase Assay kit (Promega #V9101). Reaction was initiated by mixing increasing amounts of MAPKAPK2 with 25 μM ATP in 40 mM Tris-HCl, pH 7.4, 20 mM MgCl₂, 0.1 mg/ml BSA prepared with 50 μM DTT and substrate at a final concentration of 200 µg/ml.

After a 40-minute incubation at room temperature, the reaction was terminated by addition of ADP-Glo™ Reagent, followed by a subsequent 40-minute incubation at room temperature. Kinase Detection Reagent was added, and the reaction was incubated for another 30 minutes at ambient temperature. Detection of luminescence was measured using the Luminescence Module Protocol on GloMax®-Multi Microplate Multimode Reader. The Specific Activity was calculated as follows: (Corrected activity, RLU) / [(Specific activity from ADP in RLU/pmol) * (Reaction time in min) *(Enzyme amount in μg or mg)]. Corrected RLU was calculated by subtracting the blank value from all the values. The blank was determined from a “no enzyme” sample by replacing the enzyme solution with an equal volume of Dilution Buffer IX (1x).

Instructions for Use

Thaw on ice and gently mix prior to use. DO NOT VORTEX. Perform a quick spin before opening. Aliquot into small volumes and flash freeze for long term storage. Avoid multiple freeze/thaw cycles.

Applications

Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.

Shipping Temperature

-80°C

Warnings

Avoid freeze/thaw cycles

Scientific Category

Kinase/Serine-Threonine

Regulatory Status

Research Use Only

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40116 Lot 230629

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