Nitric Oxide Synthase Assay Kit, BioAssay-Nitric Oxide Synthase Assay Kit, BioAssay代理/参数/厂家-细胞分析试剂盒-艾美捷科技有限公司

PRODUCT CENTER

细胞分析试剂盒

产品中心

当前位置：首页 > 产品中心 > 细胞生物学产品 > 细胞分析试剂盒

中文名称: Nitric Oxide Synthase Assay Kit, BioAssay

英文名字: Nitric Oxide Synthase Assay Kit, BioAssay

供应商: USBiological

产品货号: N2577-40

产品报价: ￥1/96Tests

产品说明书: 点击查看

购买方式: 银行转账、电汇、支票、现金，在线支付宝及网银支付，或直接与我们电话联系400-6800-868

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: Nitric oxide (NO) is a reactive radical that plays an important role in many key physiological functions. NO, an oxidation product of arginine by nitric oxide synthase (NOS), is involved in host defense and development, activation of regulatory proteins and direct covalent interaction with functional biomolecules.Simple, direct and non-radioactive procedures for measuring NOS are becoming popular in Research and Drug Discovery. Nitric Oxide Synthase Assay Kit involves two steps: a NOS reaction step during which NO is produced followed by an NO detection step. Since the NO generated by NOS is rapidly oxidized to nitrite and nitrate, the NO production is measured following reduction of nitrate to nitrite using an improved Griess method. The procedure is simple and the time required for sample pretreatment and assay is reduced to as short as 40 min.Key Features:Sensitive and accurate. Detection range 0.25-25 U/L in 96-well plate.Rapid and reliable. Can be completed in 40 min if reduction of NO3- to NO2- is performed at 60°C.Applications:Direct Assays: NOS activity in biological samples.Drug Discovery/Pharmacology: effects of drugs on NOS activity.Kit Contents: (100 tests in 96-well plates)Assay Buffer: 10ml Substrate: 600ul GDH: 120ulReagent A: 12ml Reagent B: 500ul Reagent C: 12mlReagent D: 600ul Reagent E: 1.5ml ZnSO4: 1mlStandard: 1ml N aOH: 1mlStorage conditions. The kit is shipped on ice. Store Assay Buffer, Substrate, Reagent D, Reagent E and GDH at -20°C. Store all other reagents at 4°C. Shelf life of three months after receipt. Precautions: reagents are for research use only. Please refer to Material Safety Data Sheet for detailed information.Procedures:Prior to assay, equilibrate all components to room temperature.Prewarm Assay Buffer to 37°C. Keep GDH on ice.Sample treatment: tissue or cell samples are homogenized in 1 x PBS (pH 7.4). Centrifuge at 10,000g or higher at 4°C. Use supernatant for NOS assay.Standard preparation: Prepare 200ul 500uM Premix by mixing 100ul 1.0mM Standard and 100ul distilled water. Dilute standards in 1.5-mL centrifuge tubes as described in the Table.No Premix+H2O Nitrite (uM)1 50ul+0ul 5002 30ul+20ul 3003 15ul+35ul 1504 0ul+50ul 0NOS Reaction: If samples will not require deproteination (i.e. purified NOS), add 20ul of each sample and standard to separate labeled eppendorf tubes. Each sample requires at least two tubes: one reaction tube and one sample blank tube. Immediately prior to starting the reaction, prepare enough NOS Working Reagent (NOS WR) for all sample reaction tubes and standards by mixing per reaction tube: 65ul Assay Buffer, 4ul Substrate, 4ul Reagent D, 10ul Reagent E and 1ul GDH. For the sample blanks, use 8ul dH2O instead of the Substrate and Reagent D. Add 80ul of the appropriate NOS WR to each tube and incubate at 37°C for 20 min. After 20 min immediately add 200ul of the NO Detection Reagent (NO DR) (see next section: NO Measurement) to each tube to kill the NOS reaction. For samples requiring deproteination which include serum, plasma, whole blood, cell culture media containing FBS, tissue or cell lysates, add 25ul of each sample and standard to separate labeled eppendorf tubes. Each sample requires at least two tubes: one reaction tube and one sample blank tube. Immediately prior to starting the reaction, prepare enough NOS WR for all sample reaction tubes and standards by mixing per reaction tube: 80ul Assay Buffer, 5ul Substrate, 5ul Reagent D, 13ul Reagent E and 1ul GDH. For the sample blanks, use 10ul dH2O instead of the Substrate and Reagent D. Add 100ul of the appropriate NOS WR to each tube and incubate at 37°C for 20 min. After 20 min immediately proceed to the deproteination step.Deproteination. Add 7ul ZnSO4 to each sample and standard tube. Vortex and then add 7ul NaOH. Vortex again and centrifuge 10 min at 14,000 rpm. Transfer 100ul of the clear supernatant to a clean tube and proceed to the NO Measurement step. NO Measurement: Immediately prior to starting the reaction, prepare enough NO Detection Reagent (NO DR) for all samples and standards by mixing per reaction tube: 100ul Reagent A, 4ul Reagent B and 100ul Reagent C. Add 200ul of the WR to each sample and standard tube and incubate for 5 min at 60°C. (Alternatively, the reaction can be run at 37°C for 60 min or RT for 150 min.) Briefly centrifuge the reaction tubes to pellet any condensation and transfer 250ul of each reaction to separate wells in a 96 well plate. Read OD at 500-570nm (peak 540 nm).Calculation:Subtract blank OD (Std 4) from the standard OD values and plot the OD against standard concentrations. Determine the slope using linear regression fitting. The NOS activity of the Sample is then calculated asODSAMPLE-ODBLANKSlopeNOS Activity=X (U/L)1tODSAMPLE and ODBLANK are optical density values of the sample and sample blank, respectively. t is the reaction time (20 min).Unit definition: one unit of NOS catalyzes the production of 1uMole of nitric oxide per minute under the assay conditions (pH 7.5 and 37°C).Materials Required, But Not Provided:Pipetting devices, eppendorf tubes, eppendorf centrifuge, clear, flat bottomed 96 well plates or cuvettes, plate reader or spectrophotometer and heat block or hot water bath (optional).General Considerations:Antioxidants and nucleophiles (e.g. b-mercaptoethanol, glutathione, dithiothreitol and cysteine) may interfere with this assay. Avoid using these compounds during sample preparation. However, if b-mercaptoethanol or dithiothreitol must be used, an equal concentration needs to be added to the standards.

产品特点: 针对Nitric Oxide Synthase 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Nitric Oxide Synthase 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。