- 中文名称
Glutathione Peroxidase, Cellular Colorimetric Assay Kit, BioAssay (c-GPx, EC 1.11.1.9)
- 英文名字
- Glutathione Peroxidase, Cellular Colorimetric Assay Kit, BioAssay (c-GPx, EC 1.11.1.9)
- 供应商
- USBiological
- 产品货号
- G8127-10
- 产品报价
- ￥1/1Kit
- 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
- Cellular glutathione peroxidase (c-GPx, EC 1.11.1.9) is a member of a family of GPx enzymes whose function is to detoxify peroxides in the cell.1 Because peroxides can decompose to form highly reactive radicals, the GPx enzymes play a critical role in protecting the cell from free radical damage, particularly lipid peroxidation. The GPx enzymes catalyze the reduction of H2O2 to water and organic peroxides (R-O-O-H) to the corresponding stable alcohols (R-OH) using glutathione (GSH) as a source of reducing equivalents: GPxR-O-O-H + 2GSH -------------> R-O-H + GSSG + H2O Equation 1With the exception of phospholipid-hydroperoxide GPx, a monomer, all of the GPx enzymes are comprised of 4 identical subunits (monomer Mr 22-23 kDa). Each subunit contains a molecule of selenocysteine in the enzyme active site. The selenocysteine is thought to participate directly in electron donation to the peroxide substrate and become oxidized in the process. The enzyme then uses glutathione as an electron donor to regenerate the reduced form of the selenocysteine.1 The GPx enzymes accept a wide variety of organic peroxides as substrates. However, with the exception of phospholipid hydroperoxide GPx and perhaps pl·GPx, the enzymes exhibit a strong preference for glutathione as a source of reducing equivalents. Phospholipid-hydroperoxide GPx (Mr 19 kDa) is the only enzyme with significant activity on esterified phospholipids and cholesterol in membranes.Principle of the Procedure:This assay is an indirect measure of the activity of c-GPx. Oxidized glutathione (GSSG), produced upon reduction of an organic peroxide by c-GPx, is recycled to its reduced state by the enzyme glutathione reductase (GR): GPxR-O-O-H + 2GSH -------------> R-O-H + GSSG + H2O Equation 2 GRGSSG + NADPH + H+ -------------> 2GSH + NADP+ Equation 3The oxidation of NADPH to NADP+ is accompanied by a decrease in absorbance at 340 nm (A340) providing a spectrophotometric means for monitoring GPx enzyme activity. The molar extinction coefficient for NADPH is6220 M-1 cm-1 at 340 nm. To assay c-GPx, a cell or tissue homogenate is added to a solution containing glutathione, glutathione reductase, and NADPH. The enzyme reaction is initiated by adding the substrate, tert-butyl hydroperoxide and the A340 is recorded. The rate of decrease in the A340 is directly proportional to the GPx activity in the sample.Samples that may be assessed using this assay include:1. Purified c-GPx: Purified enzyme can be assayed without any special preparation. However, the enzyme stock should be frozen, preferably at -70°C at 3000mU/ml or more, in a buffer containing 1mM dithiothreitol or 2- mercaptoethanol (or other thiol reducing agent), 5mM EDTA, and 1mg/ml protein (for example, bovine IgG) to preserve enzyme activity.2. Erythrocyte lysates: Recommended sample size is 0.2-0.5 mg of hemoglobin or protein (from a clarified lysate) (e.g., 70ul of 7mg/ml) added to the reaction mixture.3. Plasma or Serum: Plasma or serum are not recommended for use as samples. It has been demonstrated recently that serum albumin may have peroxidase activity.4. Cell and Tissue Homogenates: GPx activity may be assayed in most tissue or cell extracts. It is recommended that approximately 0.1mg to 1mg of protein (e.g., 70ul of 1.5 - 15mg/ml) be added to the assay in preliminary experiments to establish the range of activity expected in the particular tissue.Materials Provided (for 100 tests):G8127-10A: NADPH Reagent, 5 x 1vialBeta-nicotinamide-adenine dinucleotide phosphate (reduced), Glutathione, and Glutathione reductase. Lyophilized and when reconstituted with 7.5ml Assay Buffer, each of the 5 vials provide for 20 tests.G8127-10B: Assay Buffer (pH 7.6), 1x120mlG8127-10C: tert-Butyl Hydroperoxide, 1x2ml. Dissolved in water.Materials Required But Not Provided:1. Spectrophotometer, preferably equipped with a temperature-controlled cuvette chamber, capable of measuring the absorbance at 340nm.2. Spectrophotometric cuvettes with a 1cm path length.3. Adjustable pipettors, 10ul to 1ml with disposable pipette tips.4. 10ml glass or plastic pipettes, capable of accurately measuring 7.5ml.5. Beakers, flasks or small bottles (20-50ml capacity) for Assay Buffer and tert-Butyl Hydroperoxide.6. Deionized water7. Cellular Glutathione Peroxidase ControlStorage and Stability:Store all reagent components at 4°C.
- 产品特点
- 针对Glutathione Peroxidase, Cellular Colorimetric Assay Kit, BioAssay (c-GPx, EC 1.11.1.9)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, Cellular Colorimetric Assay Kit, BioAssay (c-GPx, EC 1.11.1.9)产品后将其置于4°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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