- 中文名称
Glutathione Peroxidase Assay Kit, BioAssay
- 英文名字
- Glutathione Peroxidase Assay Kit, BioAssay
- 供应商
- USBiological
- 产品货号
- G8127-03
- 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
- Glutathione Peroxidase (GPX, EC 1.11.1.9) represents an enzyme family with peroxidase activity whose main biological role is to protect the organism from oxidative damage. It helps prevent lipid peroxidation of cellular membranes by removing free peroxide in the cell. GPX catalyzes the following reaction with glutathione reductase (GR), GPX 2 GSH+H2O2 GS-SG+2 H2O, GR GS-SG+NADPH 2 GSH+NADP+ Simple, direct and high-throughput assays for GPX activity find wide Applications:. The improved assay directly measures NADPH consumption in the enzyme coupled reactions. The measured decrease in optical density at 340nm is directly proportional to the enzyme activity in the sample.Key Features:Sensitive and accurate. Use 10ul sample.Linear detection range 12 to 300 U/L GPX activity.Applications:Direct Assays: GPX activity in biological samples.Drug Discovery/Pharmacology: effects of drugs on GPX activity.Kit Contents: (100 Tests In 96-Well Plates)Assay Buffer: 25ml GR Enzyme: 1ml Glutathione: 240ul NADPH: 40ul H2O2 Solution: 100ul C a librator: 100ul Positive Control: 9ul Glutathione Peroxidase (GPX)Storage conditions. The kit is shipped on ice. store all components at -20°C. Shelf life of three 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.Sample Preparation:All samples should be clear and free of any turbidity or particles. Liquid samples (e.g. non-hemolyzed serum, plasma) can be assayed directly. Homogenize tissue (10 mg) and cells (106) in 200ul cold 1 x PBS and then centrifuge 10 min at 14,000 rpm to pellet any debris. Use the clear supernatant for the assay. If not assayed immediately, freeze supernatant at -80°C (stable for 1 month).Assay Procedure:1. Reagent Preparation. Equilibrate all components to room temperature. Briefly centrifuge all tubes before opening. Add 360ul dH2O to the NADPH tube (final 35mM). Add 500ul Assay Buffer to the Positive Control tube. Vortex tubes to mix. Keep these reconstituted reagent tubes on ice. Unused reagents are stable for three weeks when stored frozen at -20°C.2. Standards and Samples. Mix 12ul of the Calibrator with 188ul dH2O (equivalent to 6mM NADPH). Dilute this calibrator stock as shown in the Table below. No 6mM Calibrator+H2O Vol (ul)[Equiv. NADPH](mM)1 100ul+0ul 100 6.02 60ul+40ul 100 3.63 30ul+70ul 100 1.84 0ul+100ul 100 0Transfer 10ul standards into wells of a clear flat-bottom 96-well plate. Add 190ul Assay Buffer to all standard wells. No 6mM Calibrator+H2O Vol (ul) [Equiv. NADPH] (mM) 1 100ul+0ul 100 6.0 2 60ul+40ul 100 3.6 3 30ul+70ul 100 1.8 4 0ul+100ul 100 0 Transfer 10ul sample and 10ul reconstituted GPX Positive Control into separate wells of the 96-well plate. In addition, for each assay run, include a background control that only contains 10ul Assay Buffer. Note: (1). For unknown samples, perform several dilutions to ensure that GPX activity is within the linear range of 12 to 300 U/L. (2) The provided GPX serves as a positive control to ensure assay is working and should not be used to calculate the Sample GPX activity.3. Assay. Prepare enough Working Reagent for Sample and Control wells by mixing, for each well, 85ul Assay Buffer, 2ul Glutathione, 2ul 35mM NADPH and 8ul GR enzyme. Add 90ul Working Reagent quickly to the Sample/Control wells. Tap plate to mix. Dilute 8ul 3% H2O2 with 1992ul dH2O (final 3.5mM). Prepare enough 0.35mM H2O2 Reagent by mixing, for each Sample/Control well, 12ul 3.5mM with 108ul dH2O. Use this Reagent within one hour. With a multi-channel pipettor, add 100ul 0.35mM H2O2 Reagent to all Sample and Control wells. Tap plate quickly to mix well contents thoroughly. Immediately read OD340nm (time zero, OD0) and again at 4 min (OD4).Calculation:Use OD values at 4 min for NADPH standards. Subtract blank value (#4) from the standard values. Plot the DOD against standard concentrations and determine the slope of the standard curve. Calculate the DODs=(OD0–OD4) for the samples and DODB=(OD0–OD4) for the background control. Calculate the GPX activity of Sample, GPX Activity (U/L)=DODS-DODB Slope (mM-1) x 4 (min) °— 1000 x n The factor 1000 convertsmMoles touMoles. n is the sample dilution factor. Note: if calculated GPX activity is higher than 300 U/L, or initial OD340nm is >1.5 in sample wells, dilute sample in dH2O and repeat assay. Multiply the results by the dilution factor. Unit definition: one unit is the amount of GPX that produces 1uMole of GS-SG per min at pH 7.6 and room temperature.Materials Required, But Not Provided:Pipetting devices, centrifuge tubes, clear flat-bottom uncoated 96-well plates, plate reader capable of reading optical density at 340nm every minute, homogenizer, etc.
- 产品特点
- 针对Glutathione Peroxidase 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Glutathione Peroxidase 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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