EnzyChrom™ Adipolysis Assay Kit

I collected the medium from the culture wells. Do I need to now centifuge the medium and collect the supernatant or do I use the medium itself for the assay?

You can assay the cell culture medium directly. However, if it contains cellular debris you should first centrifuge it and then use the clear supernatant in the assay.

What is the principle of the assay?

In the process of adipolysis triglycerides are hydrolyzed by the cellular lipases into glycerol and free fatty acids. The assay measures glycerol, the end product of adipolysis, in a coupled three step enzyme reaction: 1) Glycerol kinase converts glycerol and ATP to glycerol phosphate and ADP.2) Glycerol phosphate oxidase converts glycerol phosphate and O₂ to glycerone phosphate and H₂O₂3) HRP converts ADHP and H₂O₂ to resorufin and water.For more detailed product information and questions, please feel free to Contact Us. Or for more general information regarding our assays, please refer to our General Questions.

Huttala, Outi, et al (2019). Presence of vasculature results in faster insulin response in adipocytes in vascularized adipose tissue model. ALTEX-Alternatives to animal experimentation. Assay: Adipolysis in human adipose tissue. Jocken, Johan WE, et al (2018). Short-chain fatty acids differentially affect intracellular lipolysis in a human white adipocyte model. Frontiers in endocrinology 8: 372. Assay: Adipolysis in human stem cell. Wang, Rebecca Y., et al (2017). Development of a three-dimensional adipose tissue model for studying embryonic exposures to obesogenic chemicals. Annals of biomedical engineering 45.7: 1807-1818. Assay: Adipolysis in human tissues. Zheng, Qiantao, et al (2017). Reconstitution of UCP1 using CRISPR/Cas9 in the white adipose tissue of pigs decreases fat deposition and improves thermogenic capacity. Proceedings of the National Academy of Sciences 114.45: E9474-E9482. Assay: Adipolysis in pig tissues. Huttala, Outi, et al (2016). Differentiation of human adipose stromal cells in vitro into insulin-sensitive adipocytes. Cell and tissue research 366.1: 63-74. Assay: Adipolysis in human cells. Kim, Hyo Joon, et al (2016). An apolipoprotein B100 mimotope prevents obesity in mice. Clinical Science 130.2: 105-116. Assay: Adipolysis in mouse cells. Yuan, Xiaoxue, et al (2016). Rutin ameliorates obesity through brown fat activation. The FASEB Journal 31.1: 333-345. Assay: Adipolysis in mouse cells. Chandrasekaran V., Amit A. et al (2013). Effect of Nelumbo nucifera Petal Extracts on Lipase, Adipogenesis, Adipolysis, and Central Receptors of Obesity. Evidence-Based Complementary and Alternative Medicine. 2013: 10.1155/2013/145925 Assay: Adipolysis in cell lines. Li R et al (2012). Neuropeptide Y potentiates beta-adrenergic stimulation of lipolysis in 3T3-L1 adipocytes. Regul Pept. 178(1-3):16-20. Assay: Adipolysis in mouse adipocytes.To find more recent publications, pleaseclick here.