Cell Viability Assay Kit, BioAssay(TM)

Product Overview

• Article Name: Cell Viability Assay Kit, BioAssay(TM)
• Biozol Catalog Number: USB-C2609-24
• Supplier Catalog Number: C2609-24
• Alternative Catalog Number: USB-C2609-24-500
• Manufacturer: US Biological
• Category: Kits/Assays

Product Description

The study of cell proliferation and cell viability requires the accurate quantification of the number of viable cells in a cell culture. Therefore, assays for calculating cell viability are necessary for optimizing cell culture conditions, evaluating cell growth factors and nutrients, discovering novel antibiotics and anti-cancer drugs, evaluating toxic effects of environmental pollutants and cell mediated toxicity and studying programmed cell death (apoptosis). The Cell Viability assay kit provides a convenient, sensitive, quantitative and reliable assay for determining the number of viable cells in a given culture. This homogeneous Colorimetric Assay: is based on the conversion of a tetrazolium salt MTT, a pale yellow substrate, to formazan, a purple dye. This cellular reduction reaction involves the pyridine nucleotide cofactors NADH/NADPH and is only catalyzed by living cells. The formazan product has a low aqueous solubility and is present as purple crystals. Dissolving the resulting formazan with a solubilization buffer permits the convenient quantification of product formation. The intensity of the product color, measured at 550-620nm, is directly proportional to the number of living cells in the culture. Reagents in the kit have been carefully formulated and optimized for sensitivity, assay robustness and automation. Key Features: Safe. Non-radioactive assay (cf. 3H-thymidine incorporation assay). Sensitive and accurate. As low as 950 cells can be accurately quantified. 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 and reagent transfer steps are involved. Robust and amenable to HTS. Z factors of 0.5 and above are observed. Can be readily automated with 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 screen for toxic and anticancer drugs. Kit Contents: Assay Buffer Solubilization Solution Note: Control Reagent: Saponin not supplied with kit Storage Conditions: Store the Reagent at -20C. Keep Assay Buffer and Solubilization Solution at 4 C and room temperature, respectively. 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 Cell Viability assay is based on the conversion of MTT to purple formazan by metabolically active cells. For most cells, this reducing reaction takes 3 to 5 hours. The produced formazan is solubilized and the resulting colored solution is quantified with a 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. Procedure using 96-well plate: 1. Plate and culture cells (80ul per well) in a clear bottom 96-well tissue culture plates. Typical culture medium contains DMEM, 10% fetal bovine serum, antibiotics (penicillin/streptomycin, 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 1000 to 80,000 per well. The volume can vary from 50 to 150ul, although 80ul is used in this example. In addition to the test samples, one must include control wells of culture medium containing no cells or cells treated with a toxic reagent such as 0.1% saponin. 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. A volume of 20ul in phosphate buffered saline (PBS) or culture medium is recommended for th