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Encompassing Amino Acids: full length
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: Sphingosine kinase activity was determined using D-erythro-sphingosine and ATP as substrates. Assay condition: 50 mM HEPES, pH 7.4, 150 mM NaCl, 5 mM MgCl2, 1 mM DTT, 3 µM Na orthovanadate, 0.5 mM ATP, 4 µM D-erythrosphingosine, and 5 ug/ml mouse sphingosine kinase 2 at 37°C for 30 min. ATP reduction is detected using Kinase-Glo® Luminescent Kinase Assay Platform (Promega Corporation, Madison, WI).
Description: Mouse Sphingosine kinase 2 (GenBank Accession No. NM_203280), full length with N-terminal His-tag, MW=68 kDa, expressed in a Baculovirus infected Sf9 cell expression system.
Format: Aqueous buffer solution
Formulation: 25 mM Tris-HCl, pH 8.0, 100 mM NaCl, 0.05% Tween-20, 50% glycerol, and 3 mM DTT.
Genbank: NM_203280
Host Cell Line: Sf9 cells
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal His-tag
Unit Definition: A unit of sphingosine kinase activity is defined as the amount of enzyme required to produce 1 pmol of S1P per minute at 37°C.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Zemann, B. et al., Blood.107(4):1454-8 (2006)
2. Maceyka, M. et al., J Biol Chem. 280(44):37118-29 (2005)
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