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| Product Name | Lactate Dehydrogenase Activity Colorimetric Assay Kit |
| Catalog No. | CMTR-HMM-0054 |
| Description | A colorimetric enzymatic assay for the quantification of lactate dehydrogenase activity in biological samples. The kit measures LDH-catalyzed interconversion of lactate and pyruvate coupled with NADH production or consumption, providing a spectrophotometric readout at 450 nm. |
| Intended Use | Quantitative measurement of LDH enzymatic activity in cell lysates, tissue homogenates, and biological fluids for assessment of glycolytic metabolism, tissue damage biomarker studies, and metabolic flux analysis. |
| Principle / Technology | LDH catalyzes the reversible conversion of L-lactate + NAD+ to pyruvate + NADH + H+; in this kit, LDH activity is measured in the lactate-to-pyruvate direction where NADH reduces a tetrazolium dye (WST substrate) in the presence of an electron mediator, forming a colored product with maximum absorbance at 450 nm proportional to LDH activity. |
| Detection Method | Colorimetric absorbance measurement at 450 nm using a microplate spectrophotometer, with optional kinetic reading mode |
| Sample Type | Cell lysates from cultured cells (adherent or suspension), tissue homogenates (liver, kidney, heart, skeletal muscle), serum and plasma samples, purified recombinant LDH |
| Performance Range / Specifications | Linear NADH standard curve: 0.1–10 nmol/well; LDH activity detectable over 0.01–50 mU/well range; incubation time 30 minutes at 37°C |
| Sensitivity / LOD | Detection of LDH activity from approximately 500 cells per well for cells with high glycolytic rate; lower limit approximately 0.01 mU/well purified enzyme |
| Specificity | LDH enzyme detection is specific for NAD-dependent interconversion of lactate and pyruvate; minimal interference from malate dehydrogenase or other NAD-dependent enzymes under the assay pH and substrate concentrations used |
| Reaction Conditions / Protocol | Prepare cell or tissue lysates in cold LDH Assay Buffer; add 50 μL diluted sample to microplate wells; add 50 μL LDH Reaction Mix (lactate substrate, NAD+, WST dye, and electron mediator); mix and incubate at 37°C for 30 minutes protected from light; measure absorbance at 450 nm; quantify against NADH standard curve for absolute activity values (one unit = amount of enzyme reducing 1 μmol substrate per minute at pH 7.5 at 37°C) |
| Components / Formulation | LDH Assay Buffer (Tris-HCl, pH 7.5), Lactate Substrate (1 M stock), NAD+ (lyophilized), WST-1 Solution (tetrazolium salt), Enzyme Mix (diaphorase), NADH Standard (for standard curve), Positive Control (purified rabbit muscle LDH), detailed protocol |
| Storage Conditions | All kit components at -20°C protected from light except Assay Buffer (2–8°C); reconstituted NADH standard use immediately; lactat substrate and WST solution stable for 6 months at -20°C |
| Shelf Life | 12 months from date of manufacture |
| Package Specifications | 100 assays, 400 assays (96-well format) |
| Product Form | Lyophilized and liquid reagents in individual vials |
| Quality Control | Each lot tested for linear NADH standard curve (R2 ≥0.99) and LDH positive control activity within specified range; inter-assay CV <10% |
| Key Features | Colorimetric detection compatible with routine plate readers; NADH standard curve enables absolute quantification; kit includes positive control for assay validation; applicable to all mammalian species due to conserved LDH enzyme structure |
For research use only, not for clinical use.
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