PKN3 (PRK3), GST-tag Recombinant

Catalog #

101642

$410 *

Size: 10 µg

Qty

Bulk/Custom

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Description

Recombinant human PKN3 also known as PRK3 (protein kinase N3), full length. This construct contains an N-terminal GST-tag. The recombinant protein was affinity purified and kinase active.

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Synonyms

Serine/threonine-protein kinase N3, PKNBETA

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Product Data Gallery

4-20% SDS-PAGE Coomassie Staining

Specific Activity

Product Info

Storage and Usage

Citations

Species

Human

Construct

PKN3 (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 glutatione, 0.1 mM EDTA, 0.25 mM DTT, 0.1 mM PMSF, 25% glycerol

130 kDa

Amino Acids

full length

Specific Activity

2.4 pmol/min/µg

Genbank #

NM_013355

UniProt #

Q6P5Z2

Tag(s)

N-terminal GST-tag

Storage/Stability

At least 6 months at -80°C.

Assay Conditions

Kinase activity 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.

PKN3/PRK3 activity was measured by using RSK substrate (KRRRLSSLRA) diluted in water to a final concentration of 1 mg/ml. Reaction was initiated by mixing increasing amounts of the PKN3/PRK3 with 25 µM ATP in 40 mM Tris-HCl, pH 7.4, 20 mM MgCl₂, 0.1 mg/ml BSA prepared with 250 µM DTT and substrate with a final concentration of 50 µg/ml.

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 Microplate 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

Scientific Category

Serine-Threonine Kinase

Regulatory Status

Research Use Only