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Encompassing Amino Acids: 2-587
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: Enzyme reaction is conducted in a Kinase Buffer (50 mM Tris pH 7.4, 20 mM MgCl2, 0.1 mg/ml BSA and 1 mM DTT) containing 20 µM ATP and 0.2 mg/ml MBP for 30 minutes at 30°C. Enzyme activity is measured using Kinase-Glo plus reagents (Promega).
Background: PKCι (PKC iota) is a member of the protein kinase C family of serine-threonine kinases. The amino acid sequence of PKCι showed greatest homology to PKCζ (PCKzeta), with 72% identity overall rising to 84% in the catalytic domain. Protein kinase Cι (PKCiota) has been implicated in Ras signaling and is a critical downstream effector of oncogenic Ras in the colonic epithelium. Transgenic mice expressing constitutively active PKCι in the colon are highly susceptible to carcinogen-induced colon carcinogenesis.
Description: Human PKC iota (PCKι), also known as PKClambda (PKCλ), GenBank Accession No. NM_002740, a.a. 2-587 with N-terminal GST-tag, expressed in a Baculovirus infected Sf9 cell expression system. MW= 93 kDa.
Format: Aqueous buffer solution
Formulation: 25 mM Tris-HCl, pH 8.0, 100 mM NaCl, 0.05% Tween-20, 3mM DTT and 50% glycerol.
Genbank: NM_002740
Host Cell Line: Sf9 cells
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal GST-tag
Uniprot: P41743
Unit Definition: One unit is defined as the amount of enzyme that will phosphorylate 1 pmol of Ser/Thr 7 peptide substrate per minute at pH 7.4 and 30 °C.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Zhang, J. et al., J. Biol. Chem. 279, 22118- 22123 (2004).
2. Gustrafson, W.C. et al., J. Biol. Chem. 279, 9400-9408 (2004).
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