- 中文名称
L-Lactate Assay Kit, BioAssay, Fluorometric
- 英文名字
- L-Lactate Assay Kit, BioAssay, Fluorometric
- 供应商
- USBiological
- 产品货号
- L1011-04B
- 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.Simple, direct and automation-ready procedures for measuring lactate concentration are very desirable. This Lactate Assay Kit is based on lactate dehydrogenase-catalyzed oxidation of L-Lactate, in which the formed NADH reduces a probe into a highly fluorescent product. the fluorescence intensity of this product (measured using excitation wavelength and emission wavelength of 530nm and 585 nm, respectively), is proportional to the lactate concentration in the sample.Applications:Direct Assays: Lactate in serum, plasma, urine, cell media samples and other biological samples.Key Features:1. Sensitive and accurate: Detection limit of 1uM and linearity up to 50uM L-Lactate in 96-well plate assay.2. Convenient: The procedure involves adding a single working reagent, and reading the fluorescence after 60 minutes. Room temperature assay. 3. High-throughput: Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day.Kit Components: L1011-04B1: Assay Buffer, 1x10ml L1011-04B2: NAD Solution, 1x1mlL1011-04B3: Enzyme A, 1x120ul L1011-04B4: Enzyme B, 1x120ul L1011-04B5: Probe, 1x750ulL1011-0B6: Standard, 1x1ml Storage and Stability:Store components at -20°C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vials after thawing and prior to removing the cap. Materials Required, But Not Provided:Pipeting (multi-channel) devices Black, flat bottom 96-well platesFluorescent reader capable of reading at excitation/emission wavelength of 530nm/585 nm Sample Preparation and Considerations:1. The following substances interfere and should be avoided in sample preparation: sodium azide, ascorbic acid, EDTA (>0.5mM), SDS (>0.2%), NP-40 (>1%) and Tween-20 (>1%).2. Serum and plasma samples should be diluted at 200x with dH2O. 3. Samples containing higher than 100uM pyruvate (final concentration after any dilution) require an internal standard.Protocol:1. Standard Curve: Prepare 1000ul of 40uM L-lactate Premix by mixing 2ul 20mM Standard and 998ul distilled water. For cell culture samples, prepare 1000ul of 40uM L-Lactate Premix by mixing 2ul of 20mM Standard and 998ul culture medium without serum. Dilute standard as follows:No. Premix + H2O orVol (ul)L-Lactate (uM)Medium 1 100ul + 0ul 100 402 60ul + 40ul 100 243 30ul + 70ul 100 124 0ul + 100ul 100 02. Addition of Samples and StandardsTransfer 50ul standards into wells of a black 96-well plate.Samples: Add 50ul of each sample to separate wells of a black 96-well plate.Samples requiring an internal standard will need two separate reactions:(1) Sample plus Standard and (2) Sample alone.In addition, each plate will need a Water Blank (0uM L-Lactate) reaction. For the internal standard first prepare 400ul 250uM L-Lactate standard by mixing 5ul 20mM Standard and 395ul dH2O. For the Sample plus Standard well, add 5ul 250uM L-Lactate and 45ul sample.For the Sample wells, add 5ul dH2O and 45ul sample.For the Water Blank, add 50ul dH20.2. Reagent Preparation: Spin the Enzyme tubes briefly before pipetting. For each Sample and Standard well, prepare Working Reagent by mixing 40ul Assay Buffer, 1ul Enzyme A, 1ul Enzyme B, 10ul NAD and 5ul Probe. Fresh reconstitution is recommended. 4. Reaction: Add 80ul Working Reagent per reaction well quickly. Tap plate to mix. Incubate for 60 mins. at room temperature protected from light.5. Read fluorescence under the following settings: excitation wavelength of 530nm and emission wavelength of 585nm.Calculations:Plot the L-Lactate standard curve and determine the slope. The L-Lactate concentration of the sample is calculated using the following equation:[L-Lactate] = [(Fsample -Fblank)/Slope ] x nwhere: [L-Lactate] = uM concentration of L-LactateSlope = slope of the standard curve expressed as 1/uMFsample = fluorescence intensity value of the sampleFblank = fluorescence intensity value of 0uM L-Lactate standard (Standard 4)n= dilution factor (e.g., n= 200 for serum samples)If an internal standard was needed/used, the sample L-Lactate concentration is calculated as follows:[L-Lactate] = [(Fsample - Fblank)/(Fstandard -Fsample)] x 27.8 where:[L-Lactate] = uM concentration of L-LactateFsample = fluorescence intensity value of the sampleFblank = fluorescence intensity value of Water BlankFstandard = = fluorescence intensity value of the Sample plus StandardNote: If the sample ∆F value is higher than the ∆F for 40uM L-lactate standard or greater than the ∆F for the internal standard, dilute the sample in water and repeat the assay. Multiply the results by the dilution factor.
- 产品特点
- 针对L-Lactate Assay Kit, BioAssay, Fluorometric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L-Lactate Assay Kit, BioAssay, Fluorometric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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