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Encompassing Amino Acids: full length
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: 5 mM MOPS, 2.5 mM beta-glycerophosphate, 5 mM MgCl2, 1 mM EGTA, 0.4 mM EDTA, 0.05 mM DTT, 2 mM ATP. Incubate PKCalpha with 0.2 mg/ml CREBtide peptide substrate and 50 ?M [33P]-ATP at 30°C for 15 minutes, then spot reaction on phosphocellulose paper, fix in 1% phosphoric acid, and assay with a scintillation counter.
Background: PKCα (PKC alpha) is a member of the protein kinase C (PKC) family of serine- and threonine-specific protein kinases that can be activated by calcium and the second messenger diacylglycerol. PKC-alpha has been reported to play roles in many different cellular processes, such as cell adhesion, cell transformation, cell cycle checkpoint, and cell volume control(1). PKCα has been assigned to the chromosome region 17q22-q23.2 and has been identified as a fundamental regulator of cardiac contractility and Ca(2+) handling in myocytes (2).
Description: Recombinant human PKCα (PKCalpha), full length, GenBank Accession No. NM_002737, with N-terminal GST-tag, expressed in Sf9 insect cells via a baculovirus expression system. MW= 103 kDa.
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.
Genbank: NM_002737
Host Cell Line: Sf9 cells
Purity: ≥90%
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal GST-tag
Uniprot: P17252
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Coussens, L. et al. Science 233: 859-866, 1986.
2. Braz, J. C. et al. Nature Med. 10: 248-254, 2004.
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