merck millipore,默克密理博,OB100,Adipolysis Assay Kit

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merck millipore,默克密理博,OB100,Adipolysis Assay Kit
产品名称：Adipolysis Assay Kit
产品型号：OB100
The Adipolysis Assay Kit provides the necessary reagents for differentiating adipocytes & analyzing triglyceride mobilization by glycerol release.

merck millipore,默克密理博,OB100,Adipolysis Assay Kit

产品介绍

merck millipore,默克密理博,OB100,Adipolysis Assay Kit

重要规格表

检测方法
Chromogenic

描述
产品目录编号	OB100
品牌系列	Chemicon®
商名	Chemicon
描述	Adipolysis Assay Kit
概述	Introduction 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 (RydÃ©n et al., 2002; RydÃ©n 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

产品信息
部件	Incubation Solution - (Part No. 90543) - One bottle containing 100 mL of sterile-filtered Hanks' Balanced Salt Solution. 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.
检测方法	Chromogenic

产品信息

部件

Incubation Solution - (Part No. 90543) - One bottle containing 100 mL of sterile-filtered Hanks' Balanced Salt Solution.
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.

检测方法

Chromogenic

应用
应用	The Adipolysis Assay Kit provides the necessary reagents for differentiating adipocytes & analyzing triglyceride mobilization by glycerol release.
应用说明	The 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.

应用

The Adipolysis Assay Kit provides the necessary reagents for differentiating adipocytes & analyzing triglyceride mobilization by glycerol release.

应用说明

The 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.

产品使用声明
使用声明	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.

储存和货运信息
存储条件	Note: 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.