Cell Viability Assay Kit, BioAssay(TM), High Sensitivity

Product Overview

• Article Name: Cell Viability Assay Kit, BioAssay(TM), High Sensitivity
• Biozol Catalog Number: USB-C2609-23
• Supplier Catalog Number: C2609-23
• Alternative Catalog Number: USB-C2609-23-5000
• Manufacturer: US Biological
• Category: Kits/Assays

Product Description

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 kit Storage Conditions: Store the Reagent at -20C. 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