- 中文名称
Pyruvate Assay Kit, BioAssay
- 英文名字
- Pyruvate Assay Kit, BioAssay
- 供应商
- USBiological
- 产品货号
- P9543-85
- 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
- Pyruvate is a key intermediate in cellular metabolic pathways. Pyruvate can be converted to carbohydrates via gluconeogenesis, to fatty acids or energy through acetyl-CoA, to the amino acid alanine and to ethanol. Abnormal levels of pyruvate have been linked to liver diseases and metabolic disorders.Simple, direct and automation-ready procedures for measuring pyruvate concentrations find wide applications in research and drug discovery. Pyruvate assay uses a single Working Reagent that combines pyruvate oxidase and hydrogen peroxide determination in one step. The color intensity of the reaction product at 570nm or fluorescence intensity at lem/ex=585/530nm is directly proportional to pyruvate concentration in the sample.Key Features:Sensitive and accurate. Use as little as 10ul samples. Linear detection range in 96-well plate: 2 to 500uM (17 ug/dL to 4.4 mg/dL) pyruvate for Colorimetric Assay:s and 0.2 to 50uM for Fluorometric Assay:s.Simple and convenient. The procedure involves addition of a single working reagent and incubation for 30 min at room temperature, compatible for HTS assays. Improved reagent stability. The optimized formulation has greatly enhanced the reagent and signal stability.Applications:Direct Assays: pyruvate in biological samples.Drug Discovery/Pharmacology: effects of drugs on pyruvate metabolism.Kit Contents:Enzyme Mix: 10mlDye Reagent: 120ulStandard: 400ul 25mM PyruvateStorage conditions. The kit is shipped on dry ice. Store all reagents at -20°C. Shelf life of three 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:Note: SH-group containing reagents (e.g. mercaptoethanol, DTT) may interfere with this assay and should be avoided in sample preparation.1. Equilibrate all components to room temperature. Prepare a 500uM Standard Premix by mixing 10ul of the 25mM Standard and 490ul H2O. Dilute Standard in distilled water as follows.No Premix+H2O Vol (ul) Pyruvate (uM)1 100ul+0ul 100 5002 80ul+20ul 100 4003 60ul+40ul 100 3004 40ul+60ul 100 2005 30ul+70ul 100 1506 20ul+80ul 100 1007 10ul+90ul 100 508 0ul+100ul 100 0Transfer 10ul standards and 10ul samples into separate wells of a clear flat-bottom 96-well plate.2. For each reaction well, mix 94ul Enzyme Mix and 1ul Dye Reagent in a clean tube. Transfer 90ul Working Reagent into each assay well. Tap plate to mix. Freeze unused reagents for future use.3. Incubate 30 min at room temperature. Read optical density at 570nm (550-585nm).Note: if the Sample OD is higher than the Standard OD at 500uM, dilute sample in water and repeat the assay. Multiply result by the dilution factor.Calculation:Subtract blank OD (water, #8) from the standard OD values and plot the OD against standard concentrations. Determine the slope using linear regression fitting. The pyruvate concentration of Sample is calculated as[Pyruvate] =ODSAMPLE–ODH2OSlope(uM)ODSAMPLE and ODH2O are optical density values of the sample and water.Conversions: 1mM pyruvate equals 8.7 mg/dL or 87 ppm.Fluorometric Procedure:For Fluorometric Assays, the linear detection range is 0.2 to 50uM pyruvate. Dilute the Standards prepared in Colorimetric Procedure: 1:10 in H2O.Transfer 10ul standards and 10ul samples into separate wells of a black 96-well plate.Add 90ul Working Reagent (see Colorimetric Procedure:). Tap plate to mix.Incubate 30 min at room temperature and read fluorescence at lex=530nm and lem=585nm.If assays in 384-well plate are desired, use 5ul Standards and 45ul Working Reagent. The pyruvate concentration of Sample is calculated as[Pyruvate] =FSAMPLE–FH2OSlope(uM)Materials Required, But Not Provided:Pipeting devices, centrifuge tubes, Clear flat-bottom 96-well plates, black 96-well or 384-well plates (e.g. Corning Costar) and plate reader.
- 产品特点
- 针对Pyruvate 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Pyruvate Assay Kit, BioAssay产品后将其置于-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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