- 中文名称
Phospholipase D Assay Kit, BioAssay
- 英文名字
- Phospholipase D Assay Kit, BioAssay
- 供应商
- USBiological
- 产品货号
- P4074-50P
- 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
- Phospholipase D (PLD) catalyses the hydrolysis of the phosphodiester bond of glycerophospholipids to generate phosphatidic acid and a free headgroup. Abnormalities in PLD expression have been associated with human cancers. This method provides a simple and high-throughput assay for measuring PLD activity. In this assay, PLD hydrolyzes phosphatidylcholine to choline which is determined using choline oxidase and a H2O2 specific dye. The optical density of the pink colored product at 570nm or fluorescence intensity (530/585 nm) is directly proportional to the PLD activity in the sample.Key Features:Sensitive. Use 10ul samples. Detection range: Colorimetric Assay: 0.06-10 U/L, Fluorometric Assay: 0.04-1 U/L.Simple and High-throughput: the assay involves addition of a single working reagent and can be readily adapted to high-throughput assays for drug screening.Applications:Direct Assays: phospholipase D in biological samples.Drug Discovery/Pharmacology: effects of drugs on phospholipase D metabolism.Kit Contents:Assay Buffer: 10ml Dye Reagent: 120ulEnzyme Mix: 120ul Substrate: 1.5mlCalibrator: 400ulStorage 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.Colorimetric Assay:Liquid samples can be assayed directly. Solid samples should be homogenized in a suitable enzyme buffer prior to assay.Note: SH-containing reagents (e.g. b–mercaptoethanol, dithiothreitol, > 5uM), sodium azide, EDTA, and sodium dodecyl sulfate are known to interfere in this assay and should be avoided in sample preparation. If a sample is known to contain choline, it should be removed by dialysis or membrane filtration.1. Equilibrate all components to room temperature. Briefly centrifuge the tubes before opening. Keep thawed tubes on ice during assay.2. Calibrator: mix 33ul Calibrator with 187ul dH2O (final 300uM choline). Dilute calibrator in dH2O as follows.No 300uM Premix+H2O Vol (ul) Calibrator (uM)1 100ul+0ul 100 3002 60ul+40ul 100 1803 30ul+70ul 100 904 0ul +100ul 100 0Transfer 10ul diluted standards into separate wells of a clear flatbottom 96-well plate. Samples: transfer 10ul of each sample into separate wells of the plate.3. Color reaction. Prepare enough Working Reagent by mixing, for each well, 85ul Assay Buffer, 1ul Enzyme Mix, 1ul Dye Reagent and 12ul Substrate. Add 90ul Working Reagent to each well. Tap plate to mix. Incubate at desired temperature and protect from light. At 10 and 30 min, read optical density 570nm (550-585nm).Fluorometric Assay:The Fluorometric Assay: Procedure: is similar to the Colorimetric Procedure: except that (1) 0, 9, 18 and 30uM calibrator and (2) a black 96-well plate are used. Read fluorescence intensity at lex=530 nm and l em=585 nm.Calculation:Subtract blank value (#4) from the standard values and plot the DOD or DF against standard concentrations. Determine the slope and calculate the PLD activity of Sample,[Phospholipase D] =R30–R10Slope x 20× n (U/L)R30 and R10 are optical density or fluorescence intensity readings of the Sample at 30 min and 10 min, respectively. 20 is the enzyme reaction time (30 min-10 min). n is the sample dilution factor. Note: if the calculated PLD activity of a sample is higher than 10 U/L in theColorimetric Assay: or 1 U/L in the Fluorometric Assay:, dilute sample in assay buffer and repeat the assay. Multiply result by the dilution factor.Unit definition: 1 unit of PLD catalyzes formation of 1uMole of choline per minute under the assay conditions (pH 7.4).Materials Required, But Not Provided:Pipetting devices, centrifuge tubes, clear flat-bottom uncoated 96-well plates, optical density plate reader; black flat-bottom uncoated 96-well plates, fluorescence plate reader.
- 产品特点
- 针对Phospholipase D 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Phospholipase D 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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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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