PKAcα, GST-tag Recombinant

Catalog #

40154

$410 *

Size: 10 µg

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Description

Recombinant human PKAcα (protein kinase cAMP-activated catalytic subunit alpha), full length, kinase active. The construct contains an N-terminal GST-tag. The protein kinase was affinity purified.

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Synonyms

PKAa, cAPKa, PKAcα, PRKACA, protein kinase cAMP-activated catalytic subunit α, cAMP-dependent protein kinase subunit alpha, PKA C-alpha, CAFD1, PKACA, PPNAD4

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Product Data Gallery

4-20% SDS-PAGE Coomassie Staining

Specific Activity

Product Info

Storage and Usage

Citations

Species

Human

Construct

PKAcα (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, 0.25 mM DTT, 0.1 mM EGTA, 0.1 mM EDTA, 0.1 mM PMSF, 25% glycerol.

69 kDa

Amino Acids

full length

Specific Activity

2,100 pmol/min/µg

Genbank #

NM_002730

UniProt #

P17612

Tag(s)

N-terminal GST-tag

Storage/Stability

At least 6 months at -80°C.

Assay Conditions

Kinase activity was measured using a CREBtide synthetic peptide substrate (KRREILSRRPSYR) at 1mg/ml. The protein kinase was diluted to 0.1 µg/µl in buffer containing 50ng/µl BSA and 5% glycerol. Increasing amounts of the protein kinase were mixed with 0.25 mg/ml substrate in 20 μl final volume. The blank was determined from a “no substrate” sample. The reaction was initiated by addition of 5 μl of [33P]-ATP diluted in kinase buffer: 6ml kinase buffer containing 1 mCi [33P]-ATP, 0.25 mM ATP, 25 mM MOPS, 12.5 mM β-glycero-phosphate, 25 mM MgCl₂, 5 mM EGTA, 2 mM EDTA, 0.25 mM fresh DTT, pH 7.2. 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.

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

40154 Lot 210809

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