PKAcβ, GST-tag Recombinant

Catalog #

40155

$410 *

Size: 10 µg

Qty

Bulk/Custom

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Description

Recombinant human PKAcβ (catalytic subunit C-beta of PKA), full length, with N-terminal GST-tag, expressed in Sf9 insect cells via a baculovirus expression system.

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Synonyms

PKAb; cAPKb, PKAcβ

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Product Data Gallery

4-20% SDS-PAGE Coomassie Staining

Specific Activity

Product Info

Storage and Usage

Citations

Species

Human

Construct

PKAcβ (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, 10 mM glutathione, 0.1 mM EDTA, 0.25 mM DTT, 0.1 mM PMSF, 25% glycerol.

65 kDa

Amino Acids

full length

Genbank #

NM_002731

UniProt #

P22694

Tag(s)

N-terminal GST-tag

Background

The catalytic subunit C-beta of PKA (PKAcβ) is a member of the Ser/Thr protein kinase family (the PKA catalytic subunit consist of three gene products: C-alpha, C-beta, and C-gamma) and has been assigned to human chromosome region 1p36.1 (1). PKAcβ is derived from a gene distinct from C-alpha and shows tissue-specific expression. At the amino acid level C-alpha and C-beta showed 93% homology.

References

Simard, J. et al: Assignment of the gene encoding the catalytic subunit C-beta of cAMP-dependent protein kinase to the p36 band on chromosome 1. Hum. Genet. 88: 653-657, 1992.

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.

PKAcβ kinase activity was measured by using CREBtide synthetic peptide substrate (KRREILSRRPSYR) diluted in distilled water to a final concentration of 1 mg/ml. Reaction was initiated by mixing increasing amounts of the PKAcβ 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 20 µg/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 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

Warnings

Avoid freeze/thaw cycles

Scientific Category

Kinase/Serine-Threonine

Regulatory Status

Research Use Only

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Datasheets/Protocols

40155 Lot 221018

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PKAcβ, GST-tag Recombinant

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