Citrate Synthase Assay Kit allows for the simple and convenient determination of citrate synthase activity from isolated mitochondria 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.
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.
Please read these instructions carefully before beginning this assay.For research use only. Not for human or diagnostic use.NOTE: It is recommended that gloves be worn at all time when working with isolated mitochondria and mitochondrial inhibitors.
Stability: 1 year;Storage: -20°C;This kit will perform as specified if stored as directed in the Materials Supplied section and used before the expiration date indicated on the outside of the box.
The kit will arrive as two different packages. The Citrate Synthase Activity Positive Control should be stored at 4°C. For best results, remove components and store as stated below.Citrate Synthase Activity Assay Buffer: 2 vials/15 ml, 4°C;Citrate Synthase Oxaloacetate Reagent: 1 vial/300 µg, -20°C;Citrate Synthase Acetyl Co-A Reagent: 1 vial/1 mg, -20°C;Citrate Synthase Developer Reagent: 1 vial/3 mg, -20°C;Citrate Synthase Activity Positive Control: 1 vial/10 µl, 4°C*;Half Volume 96-Well Clear Plate: 1 plate, Room temperature.