- 中文名称
Lactate Dehydrogenase (LDH) Cytotoxicity Assay Kit, BioAssay
- 英文名字
- Lactate Dehydrogenase (LDH) Cytotoxicity Assay Kit, BioAssay
- 供应商
- USBiological
- 产品货号
- L1013-98
- 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
- Lactate Dehydrogenase (LDH) is an oxidoreductase which catalyzes the interconversion of lactate and pyruvate. Cytotoxic compounds often compromise cell membrane integrity by inducing apoptosis or necrosis. LDH is a stable cytosolic enzyme that upon membrane damage is released into the cellular environment. Therefore, LDH is often measured to evaluate the presence of tissue or cell damage. The colorimetric LDH release assay is a simple and robust method to assess cytotoxic effects on cells by measuring the activity of LDH in cell culture supernatant. The assay is based on the reduction of the tetrazolium salt MTT to a formazan dye which exhibits an absorption maximum at 565 nm.Key Features:Safe. Non-radioactive assay (cf. chromium release assay).Fast. High-throughput assay using 96-well plates allows simultaneous processing tens of thousands of samples per day.Homogeneous and convenient. Mix-incubate-measure type assay. No wash steps are involved.Robust and amenable to HTS. Can be readily automated with HTS liquid handling systems.Applications:Cytotoxicity and Apoptosis: evaluation of toxic compounds, toxins, detergents, environmental pollutants etc.Drug Discovery: high-throughput screen for drug toxicity.Kit Contents: (96 wells)Substrate Buffer, 1x20ml PMS Reagent, 1x1.5mlNAD Solution, 1x1ml 10% Triton X-100, 1x1mlMTT Reagent, 1x1.5mlStorage and Stability:Store components at -20°C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Procedure:Sample Preparation: Cells should be in logarithmic growth phase for the assay. Subculture cells 2 days before the experiment. Medium containing 10% FBS is compatible with the assay. It is not necessary to subculture cells in FBS free medium.1. Plate and culture cells (200ul per well) in a 96-well tissue culture plate. Assays can be performed on either adherent cells or cells in suspension. The number of cells can varies with cell type but a range between 10,000 and 40,000 cells per well for large adherent cells and between 40,000 and 160,000 suspension cells will be appropriate for most mammalian cell types. In addition to the test samples, one must include control wells of culture medium containing no cells (Medium) and cells treated with Triton X-100 (Total Lysis).2. Add 20ul test compounds to samples wells and 20ul Triton X-100 to the Total Lysis control wells, and incubate cells for four hours or desired period of time at 37°C. It is recommended that assays be run in duplicate or triplicate.3. After the incubation remove 20ul cell-free supernatant from each well and transfer to a clear flat-bottom 96-well plate. If necessary, briefly spin the plate for 5 min at 300 x g to pellet the cells before removing the supernatant.4. Reagent Preparation: Equilibrate reagents to room temperature. The Working Reagent is prepared by mixing for each 96-well assay, 14ul MTT Solution, 8ul NAD Solution, 8ul PMS Solution and 170ul Substrate Buffer. Fresh reconstitution is recommended. Add 180ul of Working reagent per well and incubate at room temperature for 30 minutes.5. Measure OD565nm for each well in an absorbance plate reader. The suitable absorbance range for the formazan dye is between 520 and 590 nm with the maximum absorbance at 565 nm.Calculation:Cytotoxicity is calculated as the percentage of the maximum LDH release in the Total Lysis wells and in the Sample wells, as follows:Cytotoxicitiy = (ODSample–ODMedium) / (ODTotal Lysis–ODMedium) x100 (%)Where ODSample, ODMedium and ODTotal Lysis are absorbance values of the sample, the no cells control and the Triton X-100 treated control.Materials Required, But Not Provided:Pipetting devices and accessories, clear flat-bottom 96-well plates, microplate reader.Example:HL-60 leukemic cells were seeded at 120,000 cells per well immediately prior to the assay. Test compounds (Triton X-100, Saponin and Tween 20) were diluted in complete medium (RPMI1640, 10% FBS) and incubated with cells for 4 hours. Cytotoxicity of test compounds was assayed as increased release of LDH into the culture media.
- 产品特点
- 针对Lactate Dehydrogenase (LDH) Cytotoxicity 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Lactate Dehydrogenase (LDH) Cytotoxicity 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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