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細胞分析試劑盒
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中文名稱

Cell Viability Assay Kit, BioAssay, High Sensitivity

英文名字
Cell Viability Assay Kit, BioAssay, High Sensitivity
供應商
USBiological
產品貨號
C2609-23
產品報價
¥1/96Tests
產品說明書
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產品新聞
背景資料
USBiological品牌是全球有名的抗原抗體和生化試劑供應商,生產世界上種類最多的抗體,用于Western Blot、免疫沉淀、免疫熒光、免疫組化和流式細胞術等多種檢測方法。武漢艾美捷科技作為USBiological品牌中國區域總代理,是行業中少有的致力于服務客戶,幫助客戶,且擁有獨立的專業銷售團隊、技術支持團隊、市場營銷團隊、進出口報關團隊的高科技生物企業??梢詾槟峁┘皶r的咨詢響應,專業的產品和解決方案支持,穩健快捷的交貨周期,優質放心的售后服務。我們致力于為您提供有價值的產品和服務,在意您的成功!
產品描述
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.
產品特點
針對Cell Viability Assay Kit, BioAssay, High Sensitivity該產品的特點優勢,歡迎查閱官網提供的產品說明書。
保存建議
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其他
Usbiological公司是美國著名的抗體和生化試劑供應商,生產世界上種類最多的抗體,用于Western Blot、免疫沉淀、免疫熒光、免疫組化和流式細胞術等多種檢測方法。Usbiological公司現已擁有超過50,19234種抗體、抗原和生化產品,為科研用戶提供了諸多超值選擇。
注意
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