The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Specific activity is > 300 units/mg. in which one unit will convert 1.0 umole of pyruvate to L-lactate and beta-NAD per minute at pH 7.5 at 37°C.
Prepare a 1.5 ml reaction mix into a suitable container, adjust to pH 7.5 at 37°C and pre-chill on ice before use: The final concentrations are 100mM Sodium phosphate, 0.12 mM beta-nicotinamide adenine dinucleotide, reduced form, 2.3mM Pyruvate, 0.033% (w/v) bovine serum albumin.
Equilibrate to 37°C and monitor at A340nm until the value is constant using a spectrophotometer.
Add 50ul of recombinant LDHA protein in various concentrations (0.1ug, 0.5ug) in assay buffer.
Mix by inversion and record the increase at A340nm for 5 minutes.
% SDS-PAGE. ab93699 is purified by using conventional chromatography techniques.
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Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.