- 中文名称
Aspartate Transaminase Assay Kit, BioAssay
- 英文名字
- Aspartate Transaminase Assay Kit, BioAssay
- 供应商
- USBiological
- 产品货号
- A3879-62
- 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
- Aspartate Transaminase (AST), also known as serum glutamic oxaloacetic transaminase (GOT) or aspartate aminotransferase (ASAT/AAT), facilitates the conversion of aspartate and a-ketoglutarate to oxaloacetate and glutamate. There are two isoenzymes in humans: GOT1 is a cytosolic isoenzyme derived from red blood cells and heart; GOT2 is the mitochondrial isoenzyme found mainly in the liver. AST is elevated in liver and muscle diseases. It is part of diagnostic tests for liver function, myocardial infarction, acute pancreatitis, acute hemolytic anemia, severe burns, acute renal disease and trauma.Simple, direct and automation-ready procedures for measuring AST activity find wide applications in research and drug discovery. AST activity assay is based on the quantification of oxaloacetate produced by AST. In this assay, oxaloacetate and NADH are converted to malate and NAD by the enzyme malate dehydrogenase. The decrease in NADH absorbance at 340 nm is proportionate to AST activity.Key Features:Sensitive. Linear detection range: 2–100 U/L.Simple and convenient. This simple, convenient assay can be carried out in a microplate or a cuvette and takes only 10 min.Applications:Direct Assays: AST activity in serum, plasma and other biological samples.Drug Discovery/Pharmacology: effects of drugs on AST activity.Kit Contents:Assay Buffer: 24ml Cofactor: 120ulEnzyme Mix: 120ul NADH: 500ulStorage conditions. The kit is shipped on ice. Store all reagents at 20°C. Shelf life of six 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.Assay Procedure:s:Equilibrate all components to room temperature. Keep thawed enzymes on ice. Assays can be performed at 37°C or at room temperature. Prior to assay, bring the working reagents, microplate and spectrophotometer to the desired temperature.Assay is compatible with serum or plasma (heparin, EDTA). Samples should be clear and free of particles or precipitates. Hemolyzed samples should not be used.Procedure using 96-well plate1. Samples and controls. Transfer 20ul sample to each well. For each assay plate, include two wells with 20ul distilled water to be used for the NADH Standard and Blank. Keep plate at the desired temperature (e.g. 37°C).2. Prepare Working Reagent for Sample and Standard wells, by mixing for each well, 200ul Assay Buffer, 1ul Cofactor, 1ul Enzyme Mix and 4ul NADH. Warm to desired temperature (e.g. 37°C). Prepare Blank Reagent for the Blank well, by mixing 200ul Assay Buffer, 1ul Cofactor, 1ul Enzyme Mix and 4ul H2O. Warm to desired temperature (e.g. 37°C).3. Add 200ul Working Reagent to the Standard and Sample wells, and 200ul Blank Reagent to the Blank well. Immediately tap plate to mix, incubate at the desired temperature and read OD340nm at 5 min and at 10 min. Alternatively, record kinetics at 340 nm.Procedure using cuvettes1. For each assay, include one Standard and one Blank control. For each Sample and Standard, prepare Working Reagent by mixing 1000ul Assay Buffer, 5ul Cofactor, 5ul Enzyme Mix and 20ul NADH. Transfer 990ul Working Reagent to each sample cuvette and standard cuvette. Warm to desired temperature (e.g. 37°C). To Blank control cuvette, add 960ul Assay Buffer, 5ul Cofactor, 5ul Enzyme Mix and 20ul H2O. Warm to desired temperature (e.g. 37°C).2. Prewarm sample to the desired temperature. Add 100ul Sample to the Sample Cuvette. Transfer 100ul H2O to the Standard cuvette and to Blank Control cuvette, respectively.3. Mix immediately. Read OD340nm at 5 min and 10 min. Alternatively, record kinetics at 340 nm.Calculation:For each Sample, calculate the rate of NADH consumption by subtracting the OD at 10 min from the OD at 5 min (DODS). Similarly, calculate the rate (DODNADH) for the NADH standard (OD5 min-OD10min).Determine AST activity using the following equation,AST =DODS-DODNADHODSTD–ODBLK388 x (U/L)ODSTD and ODBLK are the OD340nm values of NADH Standard and Blankat 5 min, respectively. The factor 388 is derived fromFactor =4ul Vol.NADH206ul Vol.WR10mM NADH x x200ul Vol.WR220ul Vol.Totalx11 (sample dilution)5 min= 388uM/minIf the calculated AST activity is higher than 100 U/L, dilute sample in Assay Buffer and repeat assay. Multiple results by the dilution factor.Materials Required, But Not Provided:Pipeting devices and accessories. Clear bottom 96-well plates (e.g. Corning Costar) and plate reader or spectrophotometer and cuvettes for measuring OD340nm.
- 产品特点
- 针对Aspartate Transaminase 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Aspartate Transaminase 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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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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