WNK1, GST-Tag Recombinant

Catalog #

100971

$420 *

Size: 10 µg

Qty

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Description

Recombinant human WNK1 (WNK lysine deficient protein kinase 1), encompassing amino acids (181-507). This construct contains an N-terminal GST-tag. The recombinant protein was affinity purified.

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Synonyms

Protein Kinase With No Lysine 1, WNK Lysine Deficient Protein Kinase 1, Protein Phosphatase 1, Regulatory Subunit 167

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Product Data Gallery

4-20% SDS-Page Coomassie Staining

Specific Activity

Product Info

Storage and Usage

Citations

Species

Human

Construct

WNK1 (GST-181-507)

Host Species/Expression System

Sf9

Purity

≥90%

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

67 kDa

Amino Acids

181-507

Specific Activity

≥25 pmol/min/μg

Genbank #

NM_018979

UniProt #

Q9H4A3

Tag(s)

N-terminal GST-tag

Background

WNK1 is a member of the WNK subfamily of serine/threonine protein kinases that is a key regulator of blood pressure by controlling the transport of sodium and chloride ions. Mutations in WNK1 have been associated with pseudohypoaldosteronism type II and hereditary sensory neuropathy type II. WNK1 is a regulator of blood pressure and deficiency of this protein in mice lowers the blood pressure. WNK1 can regulate the Ca(2+) sensing and the subsequent Ca(2+)-dependent interactions mediated by synaptotagmin C2 domains and WNK1 exhibits additive CFTR inhibition.

Storage/Stability

At least 6 months at -80°C.

Assay Conditions

Kinase activity was measured using Myelin Basic Protein (MBP) substrate diluted in water to a final concentration of 1 mg/ml. Increasing amounts of kinase were mixed with a final concentration of 200 µg/ml peptide substrate in a buffer consisting of 5 mM MOPS pH 7.2, 5 mM MgCl2, 2.5 mM β-glycerol-phosphate, 1 mM EGTA, 0.4 mM EDTA, 50 ng/µl BSA (bovine serum albumin) and 0.25 mM fresh DTT (final concentrations). The reaction was initiated by adding 5 µl [33P]-ATP (1 µCi/sample) mixture containing 0.25 mM non-radioactive ATP. The blank was determined from a “no substrate” sample.

The reaction was incubated for 15 minutes at 30°C and 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.

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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Datasheets/Protocols

100971 Lot 220901

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