- 中文名称
p-Nitrophenyl Phosphate (pNPP, KR-pT-IRR), Phosphatase Assay Kit, BioAssay
- 英文名字
- p-Nitrophenyl Phosphate (pNPP, KR-pT-IRR), Phosphatase Assay Kit, BioAssay
- 供应商
- USBiological
- 产品货号
- N2600-37
- 产品报价
- ￥23
- 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
- Para-nitrophenyl phosphate (pNPP) is a chromogenic substrate for most phosphatases such as alkaline phosphatases, acid phosphatases, protein tyrosine phosphatases and serine/threonine phosphatases. The reaction yields para-nitrophenol, which becomes an intense yellow soluble product under alkaline conditions and can be conveniently measured at 405 nm on a spectrophotometer.Key Features:High sensitivity and wide linear range. The detection limit is generally 3ng phosphatase or below. Homogeneous and simple procedure. No wash or reagent transfer steps are involved. The assay can be completed within 30 minutes.Robust and amenable to HTS. All reagents are compatible with highthroughput liquid handling instruments.Applications:Enzyme Activity Assay and Quality Control for phosphatase production.Characterization of Kinetics of phosphatase reaction.Drug Discovery: high-throughput screen for phosphatase inhibitors.Kit Contents:Catalog # Size (assays) Reagent Assay Buffer Stop SolutionXXXX-XXX 500 280ul 25ml 25mlXXXX-XXX 1,000 550ul 50ml 50mlXXXX-XXX >10k customized customized customizedStorage conditions. The kit is shipped at ambient temperature. Store Reagent at -20°C and other components at 4°C. Shelf life: 12 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.Procedure Using 96-Well Plate1. Equilibrate all reagents to room temperature by allowing them to stand for 30 minutes at room temperature. Prepare enough pNPP Substrate by mixing per assay 0.5ul Reagent and 50ul Assay Buffer.2. Serially dilute enzyme in a proper Enzyme Buffer. Prepare enough solution for triplicate assays. Transfer 50ul of each enzyme dilution to wells of a clear, flat-bottom 96-well plate. In addition, prepare a blank control that contains 50ul Enzyme Buffer only. Initiate the reaction by adding 50ul pNPP Substrate to each well.3. Incubate for 10-30 minutes at room temperature.4. Stop the reaction by adding 50ul Stop Solution. Mix by quickly tapping the plate. Alternatively, plates can be shaken for 10 seconds on an orbital plate shaker.5. Read the absorbance of each well at 405 nm.Procedure Using 384-Well PlateThe 384-well Assay Procedure: is the same as for the 96-well plate protocol except that 25ul is mixed with 25ul pNPP Substrate. After the incubation, add 25ul Stop Solution.General Considerations:(1). Fresh reconstitution of the Reagent is recommended although the reconstituted pNPP Substrate may be stable for up to 4 weeks when stored at -20°C. (2). Assays can be performed at room temperature or at 37°C. (3). The pH of the Assay Buffer is 7.2 and is compatible with the majority of neutral phosphatases such as protein phosphatases. For an acid phosphatase, we recommend using 100mM sodium acetate (pH 5.5), 10mM MgCl2 as Enzyme Buffer. For an alkaline phosphatase, we recommend using Alkaline Phosphatase Assay Kit, BioAssay.Data Analysis:Calculate the average and standard derivations of the triplicate assays and subtract the blank values. Enzyme activity is calculated from Beer-Lambert law as follows, where e is the molar extinction coefficient (M-1 × cm-1). For p-nitrophenol, e=1.78 x 104 M-1 × cm-1. OD405nm (cm-1) is the absorbance at 405 nm divided by the light-path length (cm). V is the final assay volume, i.e., 150ul for 96-well plate assay and 75ul for 384-well plate assay.
- 产品特点
- 针对p-Nitrophenyl Phosphate (pNPP, KR-pT-IRR), Phosphatase 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p-Nitrophenyl Phosphate (pNPP, KR-pT-IRR), Phosphatase Assay Kit, BioAssay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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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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