MitoCheck® Mitochondrial (Tissue) Isolation Kit Reference: 701010-1 When measuring mitochondrial function or the effects of an unknown on mitochondrial function, it can be difficult to determine a mechanism using whole organisms or tissue. When this is the case, isolated mitochondria provide a simple and biochemically relevant experimental model. Cayman’s MitoCheck® Mitochondrial (Tissue) Isolation Kit includes a complete package of reagents and step-by-step instructions for isolating coupled mitochondria* from freshly harvested mammalian heart, liver, or kidney tissue through a process of differential centrifugation in isotonic buffers. *Coupled mitochondria are defined as mitochondria capable of phosphorylating ADP at the F1FO-ATP synthase (Complex V) through the utilization of a proton (H+) gradient, which is generated by the electron transport chain (ETC) in the presence of appropriate substrates (e.g., succinate) and in the absence of uncouplers and mitochondrial inhibitors.
MitoCheck® Citrate Synthase Activity Assay Kit Reference: 701040-96 The condensation of the dicarboxylate oxaloacetate and acetyl CoA to the tricarboxylate citrate is catalyzed by citrate synthase. It is within this reaction that carbon molecules (as acetyl CoA) obtained from pyruvate oxidation are fed into the tricarboxylic acid (TCA or citric acid) cycle. As a mitochondrial enzyme, citrate synthase is commonly used as a normalization factor for mitochondrial protein, but can also be used as a biomarker for mitochondrial content in a tissue homogenate. Cayman's MitoCheck® Citrate Synthase Activity Assay Kit allows for the simple and convenient determination of citrate synthase activity from isolated mitochondria, tissue, or cell homogenates. This assay measures the production of SH-CoA by monitoring the absorbance of Citrate Synthase Developing Reagent at 412 nm in a convenient 96-well format.