3T3-L1 Lipolysis Assay Kit, glycerol detection (reagents + cells)

This lipolysis assay kit provides the tool to study chemical compounds that may influence lipolysis in cultured adipocytes.

Principle of the Assay
Lipolytic activity is assessed by the measurement of glycerol released into the medium from triglyceride breakdown. Glycerol released to the medium is phosphorylated by adenosine triphosphate (ATP) forming glycerol-1-phosphate (G-1-P) and adenosine-5’-diphosphate (ADP) in the reaction catalyzed by glycerol kinase. G-1-P is then oxidized by glycerol phosphate oxidase to dihydroxyacetone phosphate (DAP) and hydrogen peroxide (H₂O₂). A quinoneimine dye is produced by the peroxidase catalyzed coupling of 4-aminoantipyrine (4-AAP) and sodium N-ethytl-N-(3-sulfopropyl)m-anisidine (ESPA) with H₂O₂, which shows an absorbance maximum at 540nm. The increase in absorbance at 540nm is directly proportional to glycerol concentration of the sample.