- 中文名称
Cell Viability Assay Kit, BioAssay, High Sensitivity
- 英文名字
- Cell Viability Assay Kit, BioAssay, High Sensitivity
- 供应商
- USBiological
- 产品货号
- C2609-23
- 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
- This homogeneous assay involves simply adding a single reagent to the cell culture and measuring the fluorescence intensity (excitation wavelength=530-570nm, emission wavelength=590-620nm) after an incubation step. The resazurin-based assays such as the Alamar Blue reagent, utilizes the redox dye resazurin which is not fluorescent, but upon reduction by metabolically active cells is converted into a highly fluorescent product (resorufin). Living cells can readily reduce this non-toxic reagent and the resulting increase in fluorescence intensity can be conveniently monitored using a fluorescence spectrophotometer or plate reader. Nonviable cells have no metabolic capacity and, thus, will not reduce the dye. Therefore, the fluorescence intensity observed in this assay is a true measure of the viable cells.The reagent has been optimized for maximum sensitivity, reproducibility and long shelf-life. The homogeneous cell-based assay can be performed in multi-well plates. The reagent is compatible with all culture media and with all liquid handling systems for high-throughput screening applications in 96-well and 384-well plates. applications include cell proliferation, cytotoxicity and apoptosis.Key Features:Safe. Non-radioactive assay (cf. 3H-thymidine incorporation assay). Sensitive and accurate. As low as 100 cells can be accurately quantified. Time efficient. High-throughput assay in 96-well and 384-well plates allows simultaneous processing ten of thousands of samples per day. Homogeneous and convenient. A single reagent and mix-incubate-measure type assay. No wash and reagent transfer steps are involved. Robust and amenable to HTS: Z’ factors of 0.6 to 0.9 are routinely observed in 96-well and 384-well plates. Can be readily automated on HTS liquid handling systems.Applications:Cell Proliferation: effects of cytokines, growth factor, nutrients.Cytotoxicity and Apoptosis: evaluation of toxic compounds, anti-cancer antibodies, toxins, environmental pollutants etc.Drug Discovery: high-throughput screening for anticancer drugs.Kit Contents:Reagent Note: Control Reagent: Saponin not supplied with kitStorage Conditions:Store the Reagent at -20°C. Shelf life: 12 month 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.Procedures:The assay is based on the conversion of the non-fluorescent reagent to fluorescent product by metabolically active cells. For most cells this reductive reaction takes 1 to 5 hours. The fluorescence intensity of the product is then quantified on a fluorescent microplate reader. Although most culture media contain phenol red, phenol red does not interfere with the assay. All data in Technical Notes: were obtained in culture media containing phenol red.Reagent Preparation:Important: bring reagent to room temperature before use.Procedure using 96-well plate:1. Plate and culture cells (80ul) in black 96-well tissue culture plates. Typical culture medium contains DMEM, 10% fetal bovine serum and antibiotics (penicillin/ strepto-mycin, gentamycin, etc), amino acids and other nutrients. Assays can be performed on either adherent cells or cells in suspension. The number of cells can vary from 100 to 80,000 per well. The volume can vary from 50 to 150ul, although 80ul is used in this protocol. In addition to the test samples, control wells of culture medium containing no cells or cells treated with a toxic reagent such as 0.1% saponin should be included.2. Add test compounds and controls and incubate cells for the desired period of time (typically overnight). It is recommended that assays be run in duplicate or triplicate. Compounds and controls (20ul) can be added in phosphate buffered saline (PBS) or culture medium. The Control reagent can be conveniently reconstituted with 5ml PBS (1% saponin).3. Equilibrate the Reagent to room temperature. Add 10ul (per 100ul of cell culture) of the reagent per well. The volume of the reagent can be adjusted depending on the volume of cell culture. Tap plate to mix cells with compounds. Incubate for 1 to 5 hours at 37°C.4. Measure fluorescent intensity for each well on a fluorescence plate reader. If a Molecular Devices LJL Analyst is used, use the rhodamine filter sets (530nm excitation filter, 590nm emission filter and 570nm dichroic mirror).Procedure using 384-well plate:1. Plate and culture cells (40ul) in black 384-well tissue culture plates. The number of cells can vary from 100 to 20,000 per well. The volume can vary from 25 to 60ul, although 40ul is used in this protocol. In addition to the test samples, control wells of culture medium containing no cells or cells treated with a toxic reagent such as 0.1% saponin should be included.2. Add test compounds and controls and incubate cells for the desired period of time. It is recommended that assays be run in duplicate or triplicate and that compounds be added in PBS or culture medium with a volume of 10ul.3. Equilibrate Reagent to room temperature. Add 5ul Reagent (per 50ul of cell culture). Tap plate lightly to mix Reagent with cells. Incubate for 1 to 5 hours at 37°C.4. Measure fluorescent intensity for each well on a fluorescence plate reader. If a Molecular Devices LJL Analyst is used, use the rhodamine filter sets (530nm excitation filter, 590nm emission filter and 570nm dichroic mirror).General Considerations:Incubation time. The incubation time is dependent on the cell line. Some cell lines exhibit strong metabolic activity and, thus, require shorter incubation time than less metabolically active cell lines. The incubation time can be easily determined by reading the plate multiple times e.g. every 30 minutes after adding the reagent. In general, incubation for 1 to 5 hours is sufficient. Extensive incubation (such as >18 hours) may result in non-linear fluorescence response at high cell numbers.Cell number. Generally the optimized reagent shows a broad range linear fluorescence response to the number of culturing cells. It is recommended to determine the number of cells per well that gives a highest signal:noise ratio. The optimal cell number can be easily determined by serial dilution of cells.Controls. A positive control that is either cytotoxic or promotes cell proliferation can be run although it is not required. Saponin is a cytotoxic detergent that is available for purchase. A blank control, i.e., culture medium without cells or cells containing 0.1% saponin, should be done for each assay. The blank control determines background fluorescence that must be subtracted for Data Analysis.Data Analysis:For cell proliferation or cytotoxicity assays, the activity of a test compound can be calculated as percent change in cell number as follows,Activity (%) or Cell viability (%)=100 x (Fcmpd-Fo) / (Fctrl-Fo) where Fcmpd and Fctrl are the average fluorescence intensities in the presence and absence (vehicle control) of the test compound and Fo the averaged blank control fluorescence intensity. For dose-response studies, the data are plotted against compound concentration and the EC50 for proliferation assay and IC50 for cytotoxicity assay can be determined by non-linear regression analysis using Prism or other Data Analysis: tools.Technical Notes:The assay kit has been specially optimized and formulated to provide a sensitive, convenient and robust assay for cell proliferation and cytotoxicity. Key Features: of the kits are as follows:Safe. Non-radioactive assay (cf. 3H thymidine incorporation assay). Sensitive and accurate. As low as 100 cells can be accurately quantified. Saves time. High-throughput assay in 96-well and 384-well plates allows simultaneous processing of a large of number of samples. Homogeneous and convenient. A single reagent and mix-incubate- measure type assay. No wash and reagent transfer steps are involved. Robust and amenable to HTS. Z’ factors of 0.6 to 0.9 are routinely observed in 96-well and 384-well plates. Can be readily automated on HTS liquid handling systems.
- 产品特点
- 针对Cell Viability Assay Kit, BioAssay, High Sensitivity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Cell Viability Assay Kit, BioAssay, High Sensitivity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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