A part of MilliporeSigma

OB100 | Adipolysis Assay Kit

1 kit  
Retrieving price...
Price could not be retrieved
Minimum Quantity needs to be mulitiple of
Upon Order Completion More Information
You Saved ()
Request Pricing
Limited AvailabilityLimited Availability
Limited Quantities Available
    Remaining : Will advise
      Remaining : Will advise
      Will advise
      Contact Customer Service


      Contact Customer Service

      Click To Print This Page


      Replacement Information

      Key Specifications Table

      Detection Methods
      Catalogue NumberOB100
      Brand Family Chemicon®
      Trade Name
      • Chemicon
      DescriptionAdipolysis Assay Kit

      Obesity is a significant clinical problem that contributes to life-threatening diseases such as diabetes and atherosclerosis. The identification of pathways leading to increased adipose tissue formation, and reversal of lipid stores in adipose tissue, raises the prospect of preventing or reversing obesity through pharmacological means. The process of adipogenesis, the formation of adipose tissue, has become better understood by the study of several cell types that can be induced to undergo differentiation into adipocytes. The first, and best characterized, model of adipogenesis in vitro is the 3T3-L1 cell line, a substrain of Swiss 3T3 mouse cell line (Kehinde and Green, 1974). 3T3-L1 cells propagated under normal conditions have a fibroblastic phenotype. However, when treated with a combination of dexamethasone, isobutylmethylxanthine (IBMX or MIX) and insulin, 3T3-L1 cells adopt a rounded phenotype and within 5 days begin to accumulate lipids intracellularly in the form of lipid droplets (Rubin et al., 1978).

      Adipolysis refers to lipolysis, the degradation of triglyderide stores, in differentiated adipocytes. Several compounds, including isoproterenol and tumor necrosis factor-a (TNF-a), have been shown to stimulate adipolysis in differentiated 3T3-L1 and primary human adipocytes. Isoproterenol is a nonselective agonist of the beta-adrenergic class of GPCRs, which stimulate cAMP levels in adipocytes (Robidoux et al., 2004). Subsequent activation of PKA by elevated cAMP results in phosphorylation of perilipin, which is located at the surface of the lipid droplet (Sztalryd et al., 2003). Although perilipin inhibits basal lipolysis by non-hormone sensitive lipases, phosphorylated perilipin recruits the hormone-sensitive lipase (HSL) to the surface of the lipid droplet (Zhang et al., 2003). HSL cleaves triglycerides into their constituent fatty acids and free glycerol, which can be assayed as a marker of adipolysis. Although the mechanism by which TNF-a induces adipocyte lipolysis has yet to be completely elucidated, activation of the MAPK family, downregulation of Gai, and/or downregulation of perilipin appear to play a role (Rydén et al., 2002; Rydén et al., 2004). In addition, extracellular glucose is required for the TNF-a-mediated adipocyte lipolysis (Green, et al., 2004). Glycerol generated by triglyceride breakdown is released into the extracellular space through aquaporin adipose (Kishida et al., 2000). Extracellular glycerol is easily assayed by incubation with glycerol kinase (to produce glycerol phosphate), glycerol phosphate oxidase (to produce H2O2), and horseradish peroxidase in the presence of a colorimetric substrate.

      For Research Use Only; Not for use in diagnostic procedures
      Materials Required but Not Delivered· 3T3-L1 Preadipocyte cell line (can be obtained from ATCC).

      · Media for propagation of 3T3-L1 cells - DMEM with 10% calf serum (not fetal calf serum) and antibiotics.

      · Trypsin/EDTA solution for passaging cells

      · Adipogenesis basal media - DMEM with 10% fetal calf serum and antibiotics.

      · 24-well or other size tissue culture plates for induction of adipogenesis.

      · Spectrophotometer or 96-well plate reader capable of detecting 540 nm, for the most sensitive quantitation of the Free Glycerol Assay Reagent chromophore. A spectrophotometric plate reader set at 562 nm, typically used for Bicinchoninic Acid-based protein assays, is also effective.
      Product Information
      • IBMX Solution - (Part No. 90355) - One vial containing 250 μL of 0.5 M 3-isobutyl-1-methylxanthine (IBMX) in DMSO.
      • Insulin Solution - (Part No. 90356) - One vial containing 250 μL of
      • 10 mg/mL recombinant human insulin.
      • Dexamethasone Solution - (Part No. 90357) - One vial containing
      • 100 μL of 10 mM dexamethasone in ethanol.
      • Isoproterenol Stock Solution - (Part No. 90539) - One vial containing 250 μL 10mM isoproterenol in water.
      • Sterile 30% BSA Solution - (Part No. 90540) - One bottle containing 8 mL of sterile-filtered 30% Bovine Serum Albumin in PBS.
      • Glycerol Standard Solution - (Part No. 90541) - One vial containing 0.25 mL of a 0.26 mg/mL glycerol standard.
      • Free Glycerol Assay Reagent - (Part No. 90542) - two bottles containing lyophilized Free Glycerol Assay Reagent.
      • After reconstitution in 10 mL distilled water per bottle, Free Glycerol Assay Reagent contains:
      • 0.75 mM ATP
      • 3.75 mM Magnesium salt
      • 0.188 mM 4-aminoantipyrine
      • 2.11 mM sodium-N-ethyl-N(3-sulfopropyl) m-anisidine
      • 1.25 units/mL microbial glycerol kinase
      • 2.5 units/mL microbial glycerol phosphate oxidase
      • 2.5 units/mL horseradish peroxidase
      • Buffer pH 7.0
      • 0.05% sodium azide
      • Wash Solution - (Part No. 90544) - One bottle containing 200 mL of sterile-filtered Hanks' Balanced Salt Solution.
      • Incubation Solution - (Part No. 90543) - One bottle containing 100 mL of sterile-filtered Hanks' Balanced Salt Solution.
      Detection methodChromogenic
      ApplicationThe Adipolysis Assay Kit provides the necessary reagents for differentiating adipocytes & analyzing triglyceride mobilization by glycerol release.
      Application NotesThe Chemicon Adipolysis Assay Kit provides the necessary reagents for differentiating adipocytes and analyzing triglyceride mobilization by glycerol release. The kit includes IBMX Solution, Dexamethasone Solution and Insulin Solution for addition to basal media (supplied by the user) to stimulate conversion of preadipocytes to lipid droplet-containing adipocytes. In addition, the kit supplies media optimized for application of the test substance to the differentiated adipocytes. A positive control Isoproterenol Solution is provided to stimulate high levels of adipolysis. Also included is a reagent cocktail for the coupled enzymatic analysis of free glycerol released into the medium, and a glycerol standard.

      The Chemicon Adipolysis Assay Kit is a convenient and sensitive tool for analysis of small molecules and proteins to stimulate or inhibit triglyceride breakdown in cultured adipocytes.
      Biological Information
      Physicochemical Information
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsNote: Kit components require two different storage temperatures.

      Dexamethasone Solution, IBMX Solution, Insulin Solution and Isoproterenol Stock Solution should be stored at -20ºC. Glycerol Standard Solution, Free Glycerol Assay Reagent, Sterile 30% BSA Solution, Wash Solution and Incubation Solution should be stored at 2 - 8°C.


      · Please refer to the Material Safety Data Sheet at www.chemicon.com for further precautions.
      Packaging Information
      Material Size1 kit
      Transport Information
      Supplemental Information




      Safety Data Sheet (SDS) 


      Reference overviewPub Med ID
      Obesity-induced increase in tumor necrosis factor-α leads to development of colon cancer in mice.
      Marcelo B S Flores,Guilherme Z Rocha,Danilo M Damas-Souza,Felipe Osório-Costa,Marília M Dias,Eduardo R Ropelle,Juliana A Camargo,Rita B de Carvalho,Hernandes F Carvalho,Mario J A Saad,José B C Carvalheira,Felipe Os,Mar Dias,Jos Carvalheira
      Gastroenterology 143 2012

      Show Abstract
      22677195 22677195
      The farnesoid X receptor regulates adipocyte differentiation and function by promoting peroxisome proliferator-activated receptor-gamma and interfering with the Wnt/beta-catenin pathways.
      Mouaadh Abdelkarim,Sandrine Caron,Christian Duhem,Janne Prawitt,Julie Dumont,Anthony Lucas,Emmanuel Bouchaert,Olivier Briand,John Brozek,Folkert Kuipers,Catherine Fievet,Bertrand Cariou,Bart Staels
      The Journal of biological chemistry 285 2010

      Show Abstract Full Text Article
      20851881 20851881

      User Guides

      Adipolysis Assay Kit