Malic Enzyme activity was assayed spectrophotometrically at 340nm as described in Mandela and Sauer (1975). The standard reaction mixture contained 50mM Tris-HCl, 3mM MnCl2, 5mM malate, 0.12mM NADP+, 2.5mM fumarate. Assay was performed in a Beckman spectrophotometer. The Km value is 1.5±0.6mM.
Physical Appearance
Sterile Filtered White lyophilized (freeze-dried) powder.
Formulation
Lyophilized from a 0.2μm filtered concentrated solution in PBS, pH7.4.
Endotoxin
Less than 1EU/μg of rHuME2, His-tag as determined by LAL method.
Reconstitution
We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Reconstitute in sterile distilled water or aqueous buffer containing 0.1% BSA to a concentration of 0.1-1.0mg/ml. Stock solutions should be apportioned into working aliquots and stored at ≤-20℃. Further dilutions should be made in appropriate buffered solutions.
Category
Enzymes
Background
NAD-dependent malic enzyme (ME2), mitochondrial is a protein that in humans is encoded by the ME2 gene. This gene encodes a mitochondrial NAD-dependent malic enzyme, a homotetrameric protein, which catalyzes the oxidative decarboxylation of malate to pyruvate. Three different isoforms of ME are known to be in mammalian tissues: a strictly cytosolic NADP+-dependent enzyme, an NADP+-dependent mitochondriail isoform, and a mitochondrial isoenzyme that is able to use both NAD+ and NADP+ but is more effective with NAD+. The mammalian isoforms size is about 62-64kDa. A native size of 240,000 Da proposes a tetrameric structure for the active enzyme.
