MAPKAPK3, GST-tag Recombinant

Catalog #

40117

$410 *

Size: 10 µg

Qty

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Description

Recombinant human full length MAPKAPK3 (MAP kinase-activated protein kinase 3). This construct contains an N-terminal GST-tag. The recombinant protein was affinity purified and is active.

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Synonyms

3PK; 3pK; MAPKAP3

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Product Data Gallery

4-20% SDS-PAGE Coomassie Staining

Specific Activity

Product Info

Storage and Usage

Citations

Species

Human

Construct

MAPKAPK3 (GST-Full Length)

Host Species/Expression System

Sf9 insect cells

Purity

≥90%

Format

Aqueous buffer solution

Formulation

50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.25 mM DTT, 0.1 mM EGTA, 0.1 mM EDTA, 0.1 mM PMSF, 25% glycerol

69 kDa

Amino Acids

full length

Specific Activity

553 pmol/min/µg

Genbank #

NM_004635

UniProt #

Q16644

Tag(s)

N-terminal GST-tag

Background

MAPKAPK3 is closely related to MAPKAPK2 sharing 72% nucleotide and 75% amino acid identity (1). MAPKAPK3 is activated by growth inducers and stress stimulation of cells. In vitro studies demonstrated that ERK, p38 MAP kinase and Jun N-terminal kinase were all able to phosphorylate and activate this kinase, which suggested the role of MAPKAPK3 as an integrative element of signaling in both mitogen and stress responses (2).

References

1. Sithanandam, G. et al: 3pK a new mitogen-activated protein kinase-activated protein kinase located in the small cell lung cancer tumor suppressor gene region. Molec. Cell. Biol. 16: 868-876, 1996.
2. Ludwig, S. et al: 3pK, a novel mitogen-activated protein (MAP) kinase-activated protein kinase, is targeted by three MAP kinase pathways. Molec. Cell. Biol. 16: 6687-6697, 1996.

Storage/Stability

At least 6 months at -80°C.

Assay Conditions

MAPKAPK3 activity was measured by using the HSP27tide synthetic peptide (RRLNRQLSVA-amide) at a working concentration of 1 mg/ml in distilled water, in a [33P]-ATP based assay. Reaction was initiated by mixing increasing amounts of MAPKAPK3 with 1250 pmoles of [33P]-ATP in 5 mM MOPS, pH 7.2, 2.5 mM β-glycerol-phosphate, 5 mM MgCl₂, 1 mM EGTA, 0.4 mM EDTA, 50 ng/µl BSA prepared with 50 µM DTT, 50 μM ATP and substrate at a final concentration of 200 µg/ml.

The reaction was initiated by addition of [33P]-ATP Assay Cocktail, followed by a 15-minute incubation at 30°C. The reaction was terminated by spotting the reaction mixture on phosphocellulose P81 paper, air-dry and three 10-minute washes with 1% phosphoric acid solution. Radioactivity was measured in a scintillation counter. The corrected activity (RLU) was calculated by removing the blank value for each sample. The Kinase Specific Activity was calculated as follows: RLU / [(specific activity of [33P]-ATP in cpm/pmol)*(Reaction time in min)*(Enzyme amount in µg or mg)] * [(Reaction Volume) / (Spot Volume)]. The blank was determined from a “no substrate” sample by replacing the substrate solution with an equal volume of distilled water.

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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40117 Lot 230629

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