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| | LDH Activity Assay |
| Description: | Lactate dehydrogenase activity was determined by recording the absorbance change at 340 nm produced by the oxidation of NADH. Assays were performed at 37°C. The reagent mixture contained 0.115 mM NADH, 50 mM sodium phosphate buffer, pH 7.4, sodium pyruvate (0.31 mM for LDH-C4 and LDH-B4 and 1.25 mM for LDH-A4) and the enzyme preparation, diluted with phosphate buffer, pH 7.4, to provide a ΔE340 of 0.06-0.07 per minute when the activity was assayed in a 1-cm light path. For determination of dissociation constants, the isozymes, the inhibitors (oxamate or oxamate derivatives) and the coenzyme were incubated with the buffer used in the assay for 10 min at 37°C before adding the substrate. |
| Affinity data for this assay | |
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