PKAcγ, GST-tag Recombinant

Catalog #

40156

$410 *

Size: 10 µg

Qty

Bulk/Custom

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Description

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

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Synonyms

KAPG; PKAr; cAPKr, PKAcγ

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Product Data Gallery

4-20% SDS-PAGE Coomassie Staining

Specific Activity

Product Info

Storage and Usage

Citations

Species

Human

Construct

PKAcγ (GST-Full Length)

Host Species/Expression System

Sf9 insect cells

Purity

≥85%

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_002732

UniProt #

P22612

Tag(s)

N-terminal GST-tag

Background

PKA C-gamma (PKAcγ) is a third isoform of the catalytic subunit of cAMP-dependent protein kinase. It was isolated from a human testis cDNA library and was clearly derived from a gene distinct from C-alpha and C-beta and showed tissue-specific expression. Whereas at the amino acid level C-alpha and C-beta showed 93% homology, C-gamma showed only about 80% homology to both C-alpha and C-beta (1). The PRKACG gene is intronless, contains remnants of a poly(A) tail, is flanked by direct repeats, and is co-linear with the PRKACA gene(2).

References

1. Beebe, S. J. et al: Molecular cloning of a tissue-specific protein kinase (C gamma) from human testis--representing a third isoform for the catalytic subunit of cAMP-dependent protein kinase. Molec. Endocr. 4: 465-475, 1990.
2. Reinton, N. et al: The gene encoding the C gamma catalytic subunit of cAMP-dependent protein kinase is a transcribed retroposon. Genomics 49: 290-297, 1998.

Storage/Stability

At least 6 months at -80°C.

Assay Conditions

PKAcγ kinase activity was measured CREBtide synthetic peptide (KRREILSRRPSYR) 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, 2.5 mM β-glycerol-phosphate, 5 mM MgCl₂, 1 mM EGTA and 0.4 mM EDTA with 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 0.25mM 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

40156 Lot 221018

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PKAcγ, GST-tag Recombinant

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