The Cell-Check™ Viability/Cytotoxicity Kit for Animal Cells provides a two-color fluorescence staining on both live and dead cells using two probes that measure two recognized parameters of cell viability — intracellular esterase activity and plasma membrane integrity. The kit is suitable for use with fluorescence microscopes, fluorescence multiwell plate scanners and flow cytometers and other fluorescence detection systems. The assay principles are general and applicable to most eukaryotic cell types, including adherent cells and certain tissues, but not to bacteria or yeast. It is generally faster, less expensive, safer and a more sensitive indicator of cytotoxic events than alternative methods.
Live cells are distinguished by the presence of ubiquitous intracellular esterase activity, determined by the enzymatic conversion of the virtually nonfluorescent cell-permeant calcein AM to the intensely fluorescent calcein. The polyanionic dye calcein is well retained within live cells, producing an intense uniform green fluorescence in live cells (Ex/Em ~495 nm/~520 nm). Propidium iodide (PI) enters cells with damaged membranes and undergoes a 30-fold enhancement of fluorescence upon binding to nucleic acids, thereby producing a bright red fluorescence in dead cells (Ex/Em ~528 nm/~617 nm). PI is excluded by the intact plasma membrane of live cells.
- Simplicity: Reagents are simultaneously added, no wash steps are required
- Specificity and reliability: Distinct color for live and dead cell
- Versatility: Suitable for fluorescence microscope, flow cytometer or microplate reader
|Platform:||Fluorescence Microscopy, Flow Cytometry||
|Ex/Em:||Calcein: 494/517; PI: 535/617 nm|
|Product Size:||1000 assays|
|Storage Conditions:||-20 ℃, protect from light|
|Shipping Condition:||Ice pack|
Cell viability assay
Figure 1. Live and dead Jurkat cells stained with Cell-Check™ Viability/Cytotoxicity Kit (A017). Live cells fluoresce a bright green, whereas dead cells fluoresce red.
|MSDS (PDF):||Component A||Component B|
Anal Chem (2010) 82:9506-9512
Acta Biomater (2012) –
Life Sci (2012) –
Frequently asked questions (FAQs)
I am doing a Cell-Check™ assay using Calcein, AM, for live cells and PI for dead cells. Can I fix the cells after labeling and retain the staining?
This is not recommended. Neither Calcein nor PI bind to any cellular components upon fixation. There is no guarantee that the dyes will be retained upon fixation or any subsequent wash steps. We recommend scoring for live and dead cells as soon as possible after staining.
I need to use a dead cell control for my viability assay. Do you have a protocol for killing cells for this?
Heat killing is commonly used. Place your cells in a tube in buffer and heat at 60oC for 20 minutes. You can also kill your cells by fixing them with ice cold 70% ethanol for 15 minutes. The ethanol-killed cells can then be stored at -20oC until needed, at which point you wash out the ethanol and replace with buffer.