- 中文名称
D-Lactate Assay Kit, BioAssay, Colorimetric
- 英文名字
- D-Lactate Assay Kit, BioAssay, Colorimetric
- 供应商
- USBiological
- 产品货号
- L1011-04
- 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
- Lactate is generated by lactate dehydrogenase (LDH) under hypoxic or anaerobic conditions. Monitoring lactate levels is, therefore, a good indicator of the balance between tissue oxygen demand and utilization and is useful when studying cellular and animal physiology. D-lactate is produced in only minor quantities in animals and measuring for D-lactate in animal samples is a means to determine the presence of bacterial infection.Intended Use:Simple, direct and automation-ready procedures for measuring lactate concentration are very desirable. Lactate assay kit is based on lactate dehydrogenase catalyzed oxidation of lactate, in which the formed NADH reduces a formazan (MTT) Reagent. The intensity of the product color, measured at 565nm, is proportionate to the lactate concentration in the sample.Matrix:Direct Assays: D-lactate in serum, plasma, and cell media samples.Detection Range:Detection limit of 0.05mM and linearity up to 2mM D-lactate in 96-well plate assay. For cell culture samples containing phenol red: detection limit of 0.1mM and linearity up to 1mM D-lactate in 96-well plate assay.Simple Protocol:Convenient. The procedure involves adding a single working reagent, and reading the optical density at time zero and at 20 min. Room temperature assay. High-throughput. Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day.Kit Contents: (100 tests in 96-well plates)Assay Buffer: 10ml NAD Solution: 1mlEnzyme A: 120ul MTT Solution: 1.5mlEnzyme B: 120ul Standard: 1.0ml 20mM D-lactateStorage conditions. Store all reagents at -20°C. Shelf life: 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.Protocol:Important: this assay is based on an enzyme-catalyzed kinetic reaction. Addition of Working Reagent should be quick and mixing should be brief but thorough. Use of a multi-channel pipettor is recommended.1. Standard Curve. Prepare 1000ul 2.0mM D-lactate Premix by mixing 100ul 20mM Standard and 900ul distilled water. For cell culture samples containing phenol red, prepare 1000ul 1.0mM lactate Premix by mixing 50ul 20mM Standard and 950ul culture medium without serum. Dilute standard as follows. Transfer 20ul standards into wells of a clear bottom 96-well plate.No Premix+H2O or Medium Vol (ul) D-lactate (mM)1 100ul+0ul 100 2.0 or 1.02 80ul+20ul 100 1.6 or 0.83 60ul+40ul 100 1.2 or 0.64 40ul+60ul 100 0.8 or 0.45 30ul+70ul 100 0.6 or 0.36 20ul+80ul 100 0.4 or 0.27 10ul+90ul 100 0.2 or 0.18 0ul+100ul 100 0Samples. Add 20ul sample per well in separate wells. For samples with potential endogenous enzyme activity (i.e. serum, plasma, tissue extracts), two reactions should be run: one with added Enzyme A and a No Enzyme A control. Serum and Plasma should be diluted at least 2× with dH2O prior to assay.2. Reagent Preparation. Spin the Enzyme tubes briefly before pipetting. For each Sample and Standard well, prepare Working Reagent by mixing 60ul Assay Buffer, 1ul Enzyme A, 1ul Enzyme B, 10ul NAD and 14ul MTT. Fresh reconstitution is recommended. For the No Enzyme A control, the Working Reagent includes 60ul Assay Buffer, 1ul Enzyme B, 10ul NAD and 14ul MTT.3. Reaction. Add 80ul Working Reagent per reaction well quickly. Tap plate to mix briefly and thoroughly.4. Read optical density (OD0) for time “zero” at 565 nm (520-600nm) and OD20 after a 20-min incubation at room temperature.5. Calculation:. Subtract OD0 from OD20 for the standard and sample wells. Use the DOD values to determine the sample D-lactate concentration from the standard curve. For samples requiring a No Enzyme A control, subtract the DODNoEnz value from the DODSample and use this DDOD value to determine the sample D-lactate concentration from the standard curve. Note: if the sample OD value is higher than OD for 2mM D-lactate standard, dilute sample in water and repeat the assay. Multiply the results by the dilution factor.Materials Required, But Not Provided:Pipeting (multi-channel) devices. Clear-bottom 96-well plates (e.g. Corning Costar) and plate reader.General Considerations:The following substances interfere and should be avoided in sample preparation: EDTA (>0.5mM), ascorbic acid, SDS (>0.2%), sodium azide, NP-40 (>1%) and Tween-20 (>1%).
- 产品特点
- 针对D-Lactate Assay Kit, BioAssay, Colorimetric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D-Lactate Assay Kit, BioAssay, Colorimetric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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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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