MAX-Fluor™ Fluorometric Viability Assay Kit
- Life science
- MAX-Fluor™ Fluorometric Viability Assay Kit
MAX-Fluor™ Fluorometric Cell Viability Assay Kit is a fluorescence assay that allows cell viability testing to be performed at a high level of sensitivity comparable to isotopic labeling. The specially designed CytoFluor cannot penetrate the cell membrane, but CytoFluor-AM, which is an acetoxymethyl ester bond derivative, freely passes through the cell membrane. The fluorescent CytoFluor is then separated by intracellular esterase and stays inside the cell. It is measured by fluorescence microscope and FACS (Ex/Em = 490nm/515nm). At this time, the amount of fluorescence measured is determined by the membrane permeability of living cells and the activity of esterase, thus reflecting the activity of the cells. MAX-Fluor™ can be analyzed with much higher sensitivity (30~30,000 cells) than MTZ, MTS, WST and other tetrazolium salt methods.
HEK-293 cells were grown to 24 hours in DMEM with 10% FBS and washed with PBS. EZ-Fluro™ in PBS treated to serially diluted cells in a black-walled microplate and incubated for 30 minutes at 37℃ under 5% CO₂ incubator.
1. Dilute CytoFluor-AM approximately 50-fold in PBS to the required amount. CytoFluor-AM is unstable in PBS and should be used immediately after mixing.
2. Prepare approximately 100 ㎕ of cell suspension in a 96-well plate at a density of 5,000 cells/well.
3. Incubate in a CO₂ incubator for 24 hours.
4. Add the substance to be tested at various concentrations and incubate for approximately 48 hours.
5. Remove the medium. Centrifuge in case of suspension cells and add 100 ㎕ PBS.
6. Add 10 μl of CytoFluor working solution and incubate for 10-30 minutes.
7. Analyze the fluorescence at Ex = 490 nm, Em = 515 nm.