- 中文名称
Adipolysis Assay Kit, BioAssay
- 英文名字
- Adipolysis Assay Kit, BioAssay
- 供应商
- USBiological
- 产品货号
- A0885-68A
- 产品报价
- ￥1/96Tests
- 产品说明书
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- 产品新闻
- 背景资料
- USBiological品牌是全球有名的抗原抗体和生化试剂供应商，生产世界上种类最多的抗体，用于Western Blot、免疫沉淀、免疫荧光、免疫组化和流式细胞术等多种检测方法。武汉艾美捷科技作为USBiological品牌中国区域总代理，是行业中少有的致力于服务客户，帮助客户，且拥有独立的专业销售团队、技术支持团队、市场营销团队、进出口报关团队的高科技生物企业。可以为您提供及时的咨询响应，专业的产品和解决方案支持，稳健快捷的交货周期，优质放心的售后服务。我们致力于为您提供有价值的产品和服务，在意您的成功！
- 产品描述
- Obesity is a chronic condition that develops from storage of excessive energy in the form of adipose tissue. The resulting adiposity presents a high risk factor for diseases such as type 2 diabetes, cardiovascular diseases, and cancer. ADIPOLYSIS or lipolysis is a highly regulated process in fat metabolism, in which triglycerides are broken down into glycerol and free fatty acids. Rapid, robust and accurate procedures for adipolysis quantification in cell culture are very useful in research and drug discovery. Adipolysis assay kit directly measures glycerol released during adipolysis. This homogeneous assay uses a single Working Reagent that combines glycerol kinase, glycerol phosphate oxidase and color reactions in one step. The color intensity of the reaction product at 570nm is directly proportional to glycerol concentration in the sample.Key Features:Sensitive and accurate. Use as little as 10ul samples. Linear detection range in 96-well plate: 0.92 to 100 ug/mL (10 to 1000uM) glycerol for Colorimetric Assay:s and 0.2 to 5 ug/mL for Fluorometric Assay:s.Rapid and convenient. The procedure involves addition of a single working reagent and incubation for 20 min at room temperature.Robust and amenable to HTS assays. Potential interference by testing drugs is greatly reduced at 570nm. Compatible with culture media containing phenol red. Assays can be performed in 96 or 384-well plates.Applications:Direct Assays: adipolysis (glycerol in cell culture media).Drug Discovery/Pharmacology: effects of testing drugs on adipolysis.Kit Contents: (bulk reagents available)Assay Buffer: 24ml Enzyme Mix: 500ul ATP: 250ulDye Reagent: 220ul Standard: 100ul 100mM GlycerolStorage conditions. The kit is shipped on ice. Store Assay Buffer at 4°C and other reagents at -20°C. Shelf life of 6 months after receipt. Precautions: reagents are for research use only. Normal precautions for laboratory reagents should be exercised while using the reagents. Please refer to Material Safety Data Sheet for detailed information.Colorimetric Procedure:SH-group containing reagents (e.g. mercaptoethanol, DTT) may interfere with this assay and should be avoided in sample preparation. Prior to the assay, equilibrate all components to room temperature. Keep thawed Enzyme Mix in a refrigerator or on ice during assays.1. Cell Culture. Note: Cells and testing drugs are to be provided by the customer and are not included in this reagent kit. Grow cells (e.g. preadipocytes, adipocytes) in culture plate (24-well, 96-well or 384-well). If desired, treat cells with testing drugs such as insulin, isoproterenol, and incubate for the desired time period.2. Standards and Samples. Prepare a 100ug/mL standard by mixing 10ul 100mM glycerol standard with 910ul in the same medium used for cell culture. Dilute standard in the medium as follows. Transfer 10ul standards into wells of a clear 96-well assay plate (5ul for 384-well assay plate).No 100 ug/mL STD+Medium Vol (ul) Glycerol (ug/mL)1 400ul+0ul 400 1002 300ul+200ul 500 603 150ul+350ul 500 304 0ul+500ul 500 0Collect cell culture supernatants from culture wells. Such samples should be assayed immediately or stored at -20°C. Transfer 10ul samples (5ul for 384-well assay plate) into separate wells of the assay plate.3. Enzyme Reaction. For each assay well, mix 100ul Assay Buffer, 2ul Enzyme Mix, 1ul ATP and 1ul Dye Reagent in a clean tube. Transfer 100ul Working Reagent into each assay well. Tap plate to mix. For assays in a 384-well plate, use 50ul Working Reagent per well.4. Incubate 20 min at room temperature. Read optical density at 570nm (550-585nm).Note: if the Sample OD is higher than the Standard OD at 100 ug/mL, dilute sample in water and repeat the assay. Multiply result by the dilution factor.Calculation:Subtract blank OD (#4) from the standard OD values and plot the OD against standard concentrations. Determine the slope using linear regression fitting. The glycerol concentration of Sample is calculated[Glycerol] =ODSAMPLE–ODMEDIUMSlope(ug/mL)ODSAMPLE and ODMEDIUM are optical density values of the sample andmedium (#4). Conversions: 1 ug/mL glycerol equals 10.9uM.Fluorometric Procedure:For Fluorometric Assay:s, the linear detection range is 0.2 to 5 ug/mL glycerol. Dilute Standards (#1 to # 4, see Colorimetric Procedure:) as follows: mix 10ul standard with 190ul dH2O. The glycerol concentrations are now 5.0, 3.0, 1.5 and 0 ug/mL, respectively. Cell culture supernatant: dilute by mixing 10ul cell culture sample with 190ul dH2O (dilution factor n=20). Transfer 5ul of the diluted standards and samples into separate wells of a black 96-well or 384-well plate. Add 50ul Working Reagent and tap plate to mix. Incubate 20 min at room temperature and read fluorescence at lex=530nm and lem=585nm. The glycerol concentration of Sample is calculated as[Glycerol] =FSAMPLE–FMEDIUMSlopex 20 (ug/mL)Materials Required, But Not Provided:Pipeting devices, centrifuge tubes, appropriate 96- or 384-well plates and plate reader.
- 产品特点
- 针对Adipolysis Assay Kit, BioAssay该产品的特点优势，欢迎查阅官网提供的产品说明书。
- 保存建议
- 建议收到Adipolysis Assay Kit, BioAssay产品后将其置于4°C/-20°C保存。
- 其他
- Usbiological公司是美国著名的抗体和生化试剂供应商，生产世界上种类最多的抗体，用于Western Blot、免疫沉淀、免疫荧光、免疫组化和流式细胞术等多种检测方法。Usbiological公司现已拥有超过50,19234种抗体、抗原和生化产品，为科研用户提供了诸多超值选择。
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- 注意
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该页面的中文产品信息的翻译，仅供参考。准确的产品信息请以厂家的英文说明书为准。下单前，请浏览说明书确认。
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