- 中文名称
a-Amylase Assay Kit, BioAssay, High Sensitivity
- 英文名字
- a-Amylase Assay Kit, BioAssay, High Sensitivity
- 供应商
- USBiological
- 产品货号
- A2274-14
- 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
- Amylase belongs to the family of glycoside hydrolase enzymes that break down starch into glucose molecules by acting on a-1,4- glycosidic bonds. The a-amylases (EC 3.2.1.1) cleave at random locations on the starch chain, ultimately yielding maltotriose and maltose, glucose and limit dextrin from amylose and amylopectin. In mammals, a-amylase is a major digestive enzyme. Increased enzyme levels in humans are associated with salivary trauma, mumps due to inflammation of the salivary glands, pancreatitis and renal failure.Simple, direct and automation-ready procedures for measuring amylase activity are very desirable. a-Amylase assay method involves two steps: (1). a-Amylase in the sample hydrolyzes starch and the product is rapidly converted to glucose by a-glucosidase and hydrogen peroxide by glucose oxidase; (2). hydrogen peroxide concentration is determined with a colorimetric reagent.Applications:Determination of a-amylase activity in blood, saliva, urine, grains and other agricultural samples.Key Features:Sensitive and accurate. Linear detection range 0.3 to 50 U/L a- amylase in 96-well plate assay. Convenient. The procedure involves adding a single working reagent, incubation for 15 min, followed by the detection reagent and a 20-min incubation and reading the optical density at 585 nm.Kit Contents:Assay Buffer ,pH 7.0, 1x20ml Detection Reagent, 1x20ml Glucose Standard, 1x1ml Substrate, 1x120ulEnzyme A, 1x120ulEnzyme B, 1x120ulStorage and Stability:Store Detection Reagent at 4°C. Store other components at 4°C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Materials Required, But Not Provided:Pipeting devices centrifuge tubes clear flat-bottom 96-well plates plate reader membrane filters (optional)Procedure:Reagents: Equilibrate all components to room temperature. Keep thawed Enzyme Mix in a refrigerator or on ice. The substrate may have precipitates. Prior to use, vortex tube to dissolve precipitates; gentlly swirl the Detection Reagent bottle. Sample preparation: Ideally samples are assayed fresh. When stored frozen, a-amylase is stable for one month. Ascorbic acid, heparin, EDTA, EGTA, citrate, SDS, Tris (> 8mM) and ethanol (>0.4%) interfere and should be avoided in sample preparation. If glucose is present in the sample, treat the samples as described in General Considerations. It is prudent to perform a pilot test with samples at various dilutions. Recommended dilution: serum 50-fold, saliva 2,000-fold in Assay Buffer prior to assay.1. Prepare 400uM Glucose Standard by mixing 10ul of the provided (300 mg/dL) standard with 406ul Assay Buffer. Transfer 10ul Assay Buffer, 10ul 400uM glucose, and 10ul of each sample into separate wells of a clear flat-bottom 96-well plate.2. Prepare enough Working Reagent for each well by mixing 40ul Assay Buffer, 0.5ul Substrate, 1ul Enzyme A, 1ul Enzyme B. Transfer 40ul Working Reagent to each well. Incubate for 15 min at room temperature (25°C).3. Add 150ul Detection Reagent to each well. Mix and incubate for 20 min at room temperature (25°C). 4. Read OD585nm (540-610nm) on a plate reader.Calculation:The Amylase activity is calculated asActivity =OD(sample) - OD(buffer) x400 x n (U/L) OD(standard) -OD(buffer) t(min)OD(sample), OD(standard) and OD(buffer) are optical density values of the sample, the 400uM glucose standard and Assay Buffer. t is the incubation time. t=15 min in the standard protocol. n is the dilution factor (n=50 for serum, 2000 for saliva). One unit of enzyme catalyzes the production of 1uMole of glucose per min under the assay conditions.Note: if the calculated activity is higher than 50 U/L, dilute sample in Assay Buffer and repeat assay. Multiply the results by the dilution factor.General Considerations:For samples known to contain glucose, use a membrane filter to remove glucose: 1. Load 50ul sample in a 10 kD cutoff membrane filter and add 500ul Assay Buffer. 2. Centrifuge at 14000 rpm for 30 min, check level of sample, ideally the sample level will be less than 50ul. 3. Add 500ul Assay Buffer and repeat the centrifugation. Measure final sample volume with a pipetman and calculate dilution factor n=final sample volume/50ul.
- 产品特点
- 针对a-Amylase Assay Kit, BioAssay, High Sensitivity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a-Amylase Assay Kit, BioAssay, High Sensitivity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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