Description
Key facts
Detection method:Colorimetric,
Sample types:Cell culture extracts, Tissue Extracts,
Reacts with:Mouse, Rat, Human,
Assay type:Enzyme activity,
Sensitivity:= 0.78 µg/mL,
Range:0.78 - 200 µg/mL,
Assay Platform:Microplate reader
Product details:
Malate Dehydrogenase 2 (MDH2) Activity Assay (ab119693) is used to determine mitochondrial malate dehydrogenase activity (MDH2) in a sample. The enzyme is captured within the wells of the microplate and activity is determined by following the production of NADH in the following MDH2 catalyzed reaction: malate + NAD to oxaloacetic acid + NADH (Absorbance at 450 nm). The generation of NADH is coupled to the 1:1 reduction of a reporter dye to yield a colored (yellow) reaction product whose concentration can be monitored by measuring the increase in absorbance at 450 nm. In each well, ab119693 immunocaptures only native MDH2 from the chosen sample; this removes all other enzymes, including MDH1 in cytosol.
This product allows researchers to focus on TCA cycle, studying isotype-specific malate dehydrogenase (MDH2) activity assay without the necessity of isolating mitochondria.
Other Notes
Mitochondrial malate dehydrogenase (MDH2, P40926) is a 35.5 kDa enzyme that catalyzes the conversion of malate into oxaloacetate (using NAD+) and vice versa. (EC 1.1.1.37) Several isozymes of malate dehydrogenase exist, depending on where they are localized in the cell and their specific dependence on NAD+ or NADP+ (only in chloroplasts). There are two main isoforms in eukaryotic cells. One is found in the mitochondrial matrix (MDH2), participating as a key enzyme in the citric acid cycle that catalyzes the oxidation of malate. The other is found in the cytoplasm (MDH1), assisting the malate-aspartate shuttle with exchanging reducing equivalents so that malate can pass through the mitochondrial membrane to be transformed into oxaloacetate for further cellular processes. Because malate dehydrogenase is closely tied to the citric acid cycle, regulation is highly dependent on TCA products. High malate concentrations stimulate MDH activity, and, in a converse manner, high oxaloacetate concentrations inhibit the enzyme. Enzyme activity is enhanced by acetylation.
Storage:
All components are shipped cold. Reagent dye, coupler, malate and NAD+ are shipped lyophilized. Before use rehydrate by adding 0.25 mL pure H
O to each tube and vortex each tube thoroughly to dissolve. After hydration unused amounts of these four materials should be stored at -80°C for 6 months. Store all other components at 4°C.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-+4°C, Appropriate long-term storage conditions-+4°C, Storage information-+4°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
MDH2 also known as malate dehydrogenase 2 or mitochondrial malate dehydrogenase is an enzyme with a molecular mass of approximately 35 kDa. This enzyme catalyzes the conversion of malate to oxaloacetate using NAD+ as a cofactor. It is predominantly expressed in the mitochondria where it plays an important role in cellular respiration. The enzyme enables the malate dehydrogenase reaction which is key for the functioning of the tricarboxylic acid cycle.
Biological function summary
MDH2 participates in the critical process of energy production within the cell. While it does not form a complex itself its activity is intimately connected with other enzymes in mitochondrial energy metabolism. The malate dehydrogenase assay often measures the activity of MDH2 to understand the metabolic status of cells. By facilitating the oxidation of malate MDH2 aids in maintaining the efficiency of the mitochondrial electron transport chain by regenerating NADH.
Pathways
The enzyme is essential in the tricarboxylic acid (TCA) cycle and the malate-aspartate shuttle. In the TCA cycle MDH2 collaborates with enzymes like citrate synthase and isocitrate dehydrogenase to assist in the conversion of acetyl-CoA into energy-rich molecules. The malate-aspartate shuttle on the other hand involves MDH2 working closely with aspartate transaminase to transfer reducing equivalents into the mitochondria. These pathways highlight MDH2's importance in cellular energy homeostasis.
Mutations or dysregulation in MDH2 have connections to certain metabolic conditions and cancers. For example alterations in MDH2 activity might contribute to conditions like mitochondrial myopathy altering energy metabolism. Moreover MDH2 is associated with NADH-producing enzymes whose dysregulation can support oncogenic pathways in cancer. Understanding these associations helps researchers pursue therapeutic targets that modulate MDH2 activity.
