- 中文名称
Superoxide Dismutase (SOD) Assay Kit, BioAssay
- 英文名字
- Superoxide Dismutase (SOD) Assay Kit, BioAssay
- 供应商
- USBiological
- 产品货号
- S8060-07
- 产品报价
- ￥1/96Tests
- 产品说明书
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- 产品新闻
- 背景资料
- 产品描述
- Superoxide Dismutases (SOD, EC1.15.1.1) are enzymes that catalyze the dismutation of superoxide into O2 and H2O2. They are an important antioxidant defense in all cells exposed to O2. There are three major families of superoxide dismutase: Cu/Zn, Fe/Mn, and the Ni type. Aberrant SOD activities have been linked to diseases such as amyotrophic lateral sclerosis, perinatal lethality, neural disorders and cancer. SOD assay provides a convenient colorimetric means for the quantitative determination of SOD enzyme activity in biological samples. In the assay, superoxide (O2-) is provided by xanthine oxidase (XO) catalyzed reaction. O2- reacts with a WST-1 dye to form a colored product. SOD scavenges the O2- thus less O2- is available for the chromogenic reaction. The color intensity (OD440nm) is used to determine the SOD activity in a sample.Key Features:Sensitive and accurate. Linear detection range of 0.05-3 U/mL SOD.Convenient and high-throughput. Homogeneous mix-incubate-measure type assay. No wash and reagent transfer steps are involved. Can be readily automated on HTS liquid handling systems for processing thousands of samples per day.Applications:Determination of SOD in blood, cell, tissue and other biological samples.Kit Contents:Assay Buffer: 20ml Diluent: 20mlSOD Enzyme: 120ul XO Enzyme: 120ulXanthine: 600ul WST-1: 600ulStorage conditions: store all reagents at -20°C. Shelf life of 6 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:Note: If not assayed immediately, samples can be stored at -80°C for one month. All samples can be diluted in 50mM potassium phosphate, pH 7.4.1. Tissue samples. Perfuse tissue with cold PBS to remove any red blood cells. Homogenize tissue at 5ml/g in cold lysis buffer (50mM potassium phosphate, 0.1mM EDTA, 0.5% Triton X-100). Centrifuge at 12,000g for 5 minutes at 4°C. Use supernatant for total SOD assay.2. Cell samples. Suspension cells: Centrifuge 1-2 x 106 cells at 800g for 2 minutes and discard supernatant. Wash cells with cold PBS, centrifuge, and discard the supernatant. Resuspend cells in 0.5ml of cold lysis Buffer. After 10 min on ice, centrifuge at 12,000g for 5 min. Use supernatant for total SOD assay. Adherent cells: Wash 1-2 x 106 cells cold PBS. Place dish on ice. Add 0.5ml of cold lysis buffer. After 10 min on ice, collect cells/debris with a rubber policeman. Centrifuge the cell extract at 12,000g for 5 min. Use supernatant for total SOD assay. Note: if it is desired to determine cytosolic and mitochondrial SOD activities separately, tissue/cell samples can be prepared according to Mattiazzi et al (2002). JBC 277: 29626-33.3. Blood samples: Collect serum, or plasma (heparin, citrate or EDTA) using standard protocols. The erythrocyte pellet can be lysed in 5x volume of cold dH2O; centrifuge at 12,000g for 5 min to pellet the erythrocyte membranes. Dilute serum/plasma 1:5, red cell lysate 1:100 prior to SOD assay.Prior to assay, bring all reagents to room temperature (25°C). The Xanthine reagent may appear to be turbid. Briefly vortext this tube before pipetting. Briefly centrifuge enzyme tubes, keep on ice during assay.1. Standards. Mix 8ul SOD Enzyme with 392ul Diluent to give 3U/mL SOD standard. Dilute standards as below.No 3U/mL SOD+Diluent Standard (U/mL)1 100ul+0ul 3.02 80ul+20ul 2.43 60ul+40ul 1.84 40ul+60ul 1.25 18ul+82ul 0.546 8ul+92ul 0.247 4ul+96ul 0.128 0ul+100ul 0.0Transfer 20ul SOD standards to separate wells of a clear flat-bottom 96-well plate.Transfer 20ul samples to separate wells.2. Prepare enough Working Reagent for the standard and sample wells. For each well, mix 160ul Assay Buffer, 5ul Xanthine and 5ul WST-1. Transfer 160ul Working Reagent to each well and tap plate to mix. For each well, dilute the XO Enzyme 1:20 in Diluent. Quickly add 20ul diluted XO enzyme to each assay well (use of a multi-channel pipettor is recommended). Tap plate to mix.3. Immediately read OD440nm (OD420-460nm) (ODo). Incubate for 60 min at room temperature (25°C) in the dark. Read OD440nm again (OD60).Calculation:1. For each standard and sample well, calculate DOD60=OD60-ODo.2. Calculate DDOD=DODstd8-DOD for each standard and sample where DODstd8 is the DOD for Standard # 8 (the standard with no SOD activity and highest possible absorbance).3. Plot the Standard Curve DDOD vs [SOD](U/mL). Use the DDOD for sample to determine SOD activity of sample from the standard curve.Materials Required, But Not Provided:Pipetting devices, tissue homogenizer, centrifuge and tubes, clear flatbottom 96-well plates and plate reader.
- 产品特点
- 针对Superoxide Dismutase (SOD) 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Superoxide Dismutase (SOD) 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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