Reactions were carried out with varied concentrations of substrate (10–1000 nM) and 240 pmol enzyme in Elution Buffer. Reactions containing SAN1 WT and substrate were pre-warmed to 37 °C and initiated by the addition of Start Buffer. Reactions were sampled at three time points by addition of Stop Buffer, and processed using the Nucleotide Removal kit (Qiagen). Products were quantified using Ecoscint Original scintillation liquid (National Diagnostics) and a scintillation counter and were analyzed using GraphPad Prism.
Ecoscint original
Manufactured by National Diagnostics
Ecoscint Original is a scintillation cocktail used for liquid scintillation counting. It is designed to provide efficient and reliable measurement of radioactive samples, including aqueous, organic, and biological samples.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using ecoscint original
1
Quantitative SAN1 Enzyme Kinetics
2
Radiolabeled Acetate Metabolism
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Unless specified, all chemicals were purchased from Fisher Scientific (www. fishersci.com), and solvents were at least HPLC grade. [ 14 C]acetate sodium salt 50 mCi/mmol was from American Radiolabeled Chemicals, Inc. Glass thinlayer chromatography (TLC) plates (hard-layer 250 mm, 20 3 20 cm) were from Analtech. The liquid scintillation fluid was EcoScint Original (National Diagnostics). Lipase from Rhizomucor miehei and phospholipase A2 from Apis mellifera were purchased from Sigma-Aldrich.
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