Description
Key facts
Detection method:Colorimetric,
Sample types:Cell culture supernatant, Adherent cells,
Assay type:Quantitative,
Assay Platform:Microplate reader
Product details:
Abcam's Adipolysis Assay Kit (ab133115) provides a convenient tool for studying the adipolysis of triglycerides in differentiated 3T3-L1 cells, using a common adipogenesis induction procedure. This kit will allow investigators to screen compounds involved in lipid storage and metabolism. Isoproterenol is included in the kit as a positive control for screening pharmaceuticals that regulate FFA release from adipocytes.
Triglycerides stored in these lipid droplets can be stimulated by known factors or compounds to release FFA and glycerol. The latter can be measured by incubation with glycerol kinase, glycerol phosphate oxidase, and horseradish peroxidase in the presence of a colorimetric substrate to generate a chromophore detectable at 540 nm. The amount of glycerol released into the medium is proportional to the level of triglyceride storage and/or degree of adipolysis.
Obesity is a risk factor in many of the major chronic diseases such as diabetes mellitus, and cancer. The ability to regulate the number of fat cells and/or the size of fat cells is a key in the development and physiology of obesity and also in the origin of chronic disease. Adipose tissue plays an important role in energy homeostasis. Mammals have both brown adipose tissue (BAT), which utilizes lipids to generate heat in a process known as thermogenesis, and white adipose tissue (WAT) which stores excess energy as triglycerides in lipid droplets. Triglycerides can be broken down into free fatty acids (FFA) and glycerol, through a process of adipolysis (also referred as lipolysis).
Adipolysis
is a highly regulated process which allows appropriate delivery of FFA to meet energy needs. Appropriate FFA levels are important for optimal human health but excess FFAs are generally associated with development of insulin resistance, hypertriglyceridemia, reduced hepatic insulin clearance, and impaired β-cell insulin secretion. Adipolysis is under tight hormonal control.
3T3-L1 cells are a well-characterized model for studying lipid metabolism, including the uptake and release of fatty acids from fat cells. This model shares similar morphology, gene expression, and metabolism with adipocytes
in vivo
. During terminal differentiation, the fibroblast-like preadipocytes undergo a series of morphological and biochemical changes to eventually accumulate lipid droplets.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Adipolysis refers to the breakdown of lipids involving a process where triglycerides are hydrolyzed into free fatty acids and glycerol. In this process enzymes like hormone-sensitive lipase (HSL) and adipose triglyceride lipase (ATGL) also known as PNPLA2 play significant roles. HSL has a mass of approximately 84 kDa. These enzymes are widely expressed in adipose tissue which is the primary site of lipid storage in the body. Adipolysis is an important metabolic function critical for energy mobilization during fasting or increased energy demand states.
Biological function summary
The process of breaking down triglycerides into fatty acids and glycerol provides energy for cells when glucose levels are low. This process does not occur in isolation; it involves a complex interplay of hormonal signals. For instance catecholamines like epinephrine bind to β-adrenergic receptors on adipocytes triggering a cascade that leads to the activation of lipolytic enzymes. Moreover insulin modulates adipolysis by inhibiting these enzymes maintaining balance in lipid storage and usage.
Pathways
Adipolysis integrates closely with the catabolism of fatty acids a primary component of the energy-providing β-oxidation pathway. The released free fatty acids from adipose tissue enter the bloodstream and reach the liver and muscle cells where they undergo oxidation for energy production. Additionally the signaling pathway involving the AMP-activated protein kinase (AMPK) is relevant as AMPK activation promotes energy expenditure and fat breakdown linking adipolysis to metabolic rate regulation.
The dysregulation of adipolysis can contribute to metabolic syndromes like obesity and type 2 diabetes. Excessive lipolysis may lead to increased circulating free fatty acids causing insulin resistance and steatosis where inappropriate lipid accumulation occurs in liver cells. Moreover proteins such as perilipin which acts as a protective layer on lipid droplets often exhibit altered expression in these conditions affecting the regulation of adipolytic activity. Understanding these mechanisms is key in developing therapeutic strategies for related metabolic disorders.
