Malate Dehydrogenase Assay
Malate dehydrogenase (MDH) (EC 126.96.36.199) is an enzyme which reversibly catalyzes the oxidation of L-malate to oxaloacetate in the presence of NAD. There are 2 isoforms in eukaryotic cells: MDH1 and MDH2. MDH1 found in the cytoplasm and plays a key part in the malate-aspartate shuttle for transporting malate into the mitochondria. MDH2 is a mitochondrial enzyme which participates in the TCA cycle that reversibly converts L-malate into oxaloacetate. Higher MDH activities are found in some neurodegenerative diseases such as Alzheimer's disease.Assay Genie's non-radioactive, colorimetric Malate Dehydrogenase assay is based on the reduction of the tetrazolium salt MTT in a NADH-coupled enzymatic reaction to a reduced form of MTT which exhibits an absorption maximum at 565 nm. The increase in absorbance at 565 nm is proportional to the enzyme activity.
ApplicationsFor quantitative determination of L-malate dehydrogenase enzyme activity and drug effects on malate dehydrogenase metabolism.
- Fast and sensitive. Linear detection range (20 uL sample): 0.5 to 65 U/L for 20 min reaction at 37°C.
- Convenient and high-throughput. Homogeneous "mix-incubate-measure" type assay. Can be readily automated on HTS liquid handling systems for processing thousands of samples per day.
Malate Dehydrogenase Assay Information
|Kit Includes:||Assay Buffer: 10 mL, Enzyme A: 120 uL, NAD/MTT: 1 mL, Enzyme B: 120 uL, Substrate: 600 uL, Calibrator: 1.5 mL|
|Kit Requires:||Pipetting devices and accessories (e.g. multi-channel pipettor), clear flat- bottom 96-well plates, centrifuge tubes and plate reader.|
|Method of Detection:||OD565nm|
|Detection Limit:||0.5 U/L|
|Samples:||Biological samples (e.g. plasma, serum, erythrocytes, tissue and culture media).|
|Protocol Length:||30 min|
|Storage:||Store all components at -20°C upon receiving|
|Shelf Life:||6 months|