NUAK2 (K81R), Inactive, GST-Tag Recombinant

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Catalog #

101474

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Description

Recombinant human NUAK2 (SNF1—like kinase 2) full length protein containing an N-terminal GST-tag. This construct contains mutation K81R. This recombinant protein was affinity purified and is not kinase active.

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Synonyms

NUAK family SNF1-like kinase 2, Omphalocele kinase 2, SNF1/AMP kinase-related kinase, SNARK, OMPHK2

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Product Data Gallery

4-20% SDS-Page Coomassie Staining

Activity

Product Info

Storage and Usage

Citations

Species

Human

Construct

NUAK2 (K81R) (GST-Full Length)

Mutation

K81R

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.

110 kDa

Amino Acids

Full Length

Genbank #

NM_030952

UniProt #

Q9H093

Tag(s)

N-terminal GST-tag

Storage/Stability

At least 6 months at -80°C.

Assay Conditions

Kinase activity was measured using a CHKtide peptide substrate (KKKVSRSGLYRSPSMPENLNRPR) diluted in distilled water to a final concentration of 1 mg/ml.
Increasing amounts of kinase were mixed with CHKtide peptide substrate with a final concentration of 200 µg/ml in a buffer containing 5 mM MOPS, pH 7.2, 2.5 mM β-glycerol-phosphate, 5 mM MgCl2, 1 mM EGTA, 0.4 mM EDTA and 0.05 mM fresh DTT to a final volume of 20 µl. The reaction was initiated by addition of 5 μl of [33P]-ATP diluted in kinase buffer: 6 ml kinase buffer containing 1 mCi [33P]-ATP, 0.25 mM ATP, 25 mM MOPS, pH 7.2, 12.5 mM β-glycero-phosphate, 25 mM MgCl2, 5 mM EGTA, 2 mM EDTA, and 0.25 mM fresh DTT.
After incubating for 30°C for 15 minutes, the reaction was terminated by spotting 20 μl of the mixture onto phosphocellulose paper strips that were fixed in 1% phosphoric acid and washed three times. Radioactivity was determined using a scintillation counter. The blank was determined from a “no substrate” sample.

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.

Warnings

Avoid freeze/thaw cycles.

Scientific Category

Serine-Threonine Kinase

Regulatory Status

Research Use Only