MSK1, GST-Tag Recombinant

Catalog #

40131

$410 *

Size: 10 µg

Qty

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Description

Recombinant human MSK1, full length, with N-terminal GST-tag, expressed by Sf9 insect cells via a baculovirus expression system.

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Synonyms

RPS6KA5, RLPK, MSPK1, MGC1911

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Product Data Gallery

4-20% SDS-PAGE Coomassie Staining

Specific Activity

Product Info

Storage and Usage

Citations

Species

Human

Construct

MSK1 (GST-Full Length)

Host Species/Expression System

Sf9

Purity

≥70%

Format

Aqueous buffer solution

Formulation

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

120 kDa

Amino Acids

Full Length

Specific Activity

9.9 pmol/min/µg

Genbank #

NM_004755

UniProt #

O75582

Tag(s)

N-terminal GST-tag

Background

MSK1 or mitogen- and stress-activated protein kinase-1 contains 2 protein kinase domains and shares 43% protein sequence identity with the MAPKAPK1 isoforms. Northern blot analysis shows that MSK1 is expressed in all tissues with the highest level of expression in brain, muscle, and placenta. Immunoelectron microscopy localized MSK1 to the nucleus. MSK1 is activated in vitro and in vivo by either ERK or SAPK2 proteins. MSK1 rather than MAPKAP-K1 or MAPKAP-K2/K3, mediates activation of the cAMP response element-binding protein and activating transcription factor-1 by either growth factors or stress signals. By radiation hybrid analysis, the RPS6KA5 gene has been mapped to chromosome 14q31-q32.

References

1. Deak, M. et al: Mitogen- and stress-activated protein kinase-1 (MSK1) is directly activated by MAPK and SAPK2/p38, and may mediate activation of CREB. EMBO J. 17: 4426-4441, 1998.
2. Jiang, C. et al: Assignment of a member of the ribosomal protein S6 kinase family, RPS6KA5, to human chromosome 14q31-q32.1 by radiation hybrid mapping. Cytogenet. Cell Genet. 87: 261-261, 1999.

Storage/Stability

At least 6 months at -80°C.

Assay Conditions

The kinase activity of the complex was measured using the ADP-Glo™ Kinase Assay kit (Promega; Cat#V9101) which quantifies the amount of ADP produced. The ADP-Glo™ Reagent is added to terminate the reaction and deplete the remaining ATP. The Kinase Detection Reagent is then added to convert ADP to ATP and to measure the newly synthesized ATP using a luciferase reaction.

Kinase activity was measured RSK-sub peptide substrate (KRRRLSSLRA) diluted in water to a final concentration of 1 mg/ml. Reaction was initiated by mixing increasing amounts of the MSK1 with 0.125 µM ATP in 40 mM Tris-HCl, pH 7.4, 20 mM MgCl₂, 0.1 mg/ml BSA, 250 µM DTT with the 1 mg/ml substrate.

After a 40-minute incubation at 37°C, the reaction was terminated by addition of the AMP-Glo™ Reagent followed by a subsequent 40 minute incubation at room temperature. Kinase Detection Reagent was then added and incubated for another 30 minutes. Detection of luminescence was measured using the Luminescence Module Protocol on GloMax®-Multi Micorplate reader. The corrected activity (RLU) was calculated by removing the blank value for each sample divided by the (specific activity of ADP in RLU/pmol)*(Reaction time in min)*(Enzyme amount in µg or mg). The blank was determined from a “no kinase” sample by replacing the enzyme working solution with an equal volume of Kinase 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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40131 Lot 220901

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