Bovine Malate dehydrogenase, mitochondrial (MDH2) ELISA Kit

The Assay Genie Bovine MDH2 Sandwich ELISA Kit uses a colorimetric readout on a 96-well microplate, pairing a pre-coated capture antibody with a matched detection antibody raised against mitochondrial malate dehydrogenase. The assay is configured for in vitro research use only and accepts bovine serum and plasma samples, supporting quantitative work in mitochondrial enzyme biology, TCA cycle characterization, and biomarker profiling of metabolic or hepatic models. Reconstituted standards and detection reagent should be handled on ice and stored at 2-8 °C, while unused strips from the microplate may be returned to the foil pouch with desiccant and kept refrigerated for short-term use. Each kit is supplied with a pre-coated 96-well plate, lyophilized recombinant MDH2 standards, biotinylated detection antibody, streptavidin-HRP conjugate, wash buffer, substrate solution, and stop solution, sufficient for up to 40 samples measured in duplicate. Typical intra-assay and inter-assay precision allow reliable measurement across the standard curve, and the assay is intended to be read on a standard microplate reader at 450 nm with a reference wavelength.

Mitochondrial malate dehydrogenase (MDH2) catalyzes the reversible NAD+/NADH-dependent interconversion of malate and oxaloacetate inside the mitochondrial matrix, making it a frequently measured indicator of TCA cycle flux and mitochondrial integrity in bovine metabolic research. The Assay Genie sandwich ELISA detects the native dimeric protein, which complements enzymatic activity assays that can be confounded by the cytosolic MDH1 isoform and by lactate dehydrogenase cross-reactivity in crude tissue homogenates.

For sample handling, serum and plasma should be clarified by centrifugation at approximately 1000 x g for 10 minutes, then aliquoted and stored at -80 °C if not assayed within 24 hours; avoid more than two freeze-thaw cycles to preserve antibody-recognized epitopes. Reconstituted standards and working detection antibody remain stable for roughly 4 weeks at 2-8 °C, while the lyophilized stock components are shelf-stable for several months under the recommended storage conditions. The standard curve typically spans the low ng/mL range, read on a standard 450 nm microplate reader with an optional 630 nm reference filter to subtract plate-bottom noise.

Reported applications include tracking hepatic mitochondrial enzyme release in nutritional and metabolic studies, profiling skeletal muscle biopsies, comparing mitochondrial content in high- versus low-yield dairy cattle, and screening culture supernatants from bovine cell models under oxidative stress. Each 96-well plate accommodates an eight-point standard curve run in duplicate plus up to 40 unknown samples measured in duplicate, and the format supports batch processing for longitudinal or cohort-based studies.