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Encompassing Amino Acids: 1-537(end)
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: CERK activity was measured at 37°C for 25 minutes in a buffer (20 mM HEPES pH 7.2, 80 mM KCl, 3 mM CaCl2, 10 mM MgCl2, 2 mM DTT, 5 µM ATP) using a beta-Octylglucoside mixed micellar assay with 100 µM Ceramide (BPS# 40618). ATP reduction was detected using Kinase-Glo® Luminescent Kinase Assay Platform (Promega, Madison, WI).
Description: Human ceramide kinase (CERK), GenBank Accession No. NM_022766, a.a. 1-537(end) with C-terminal FLAG-tag, MW= 60 kDa, expressed in a baculovirus expression system.
Format: Aqueous buffer solution
Formulation: 40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 40 ug/ml FLAG peptide, 20% glycerol and 3 mM DTT.
Genbank: NM_022766
Host Cell Line: Sf9 cells
Storage Stability: At least 6 months at -80°C.
Tags: C-terminal FLAG-tag
Uniprot: Q8TCT0
Unit Definition: A unit of ceramide kinase activity is defined as the amount of enzyme required to produce 1 pmole of ceramide-1-phosphate/min at 37°C.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Mitra,P. et al., FEBS Lett. 581 (4), 735-740 (2007)
2. Kim,T.J. et al., Biochem. Biophys. Res. Commun. 342 (2), 611-617 (2006)