 |
 |
 |
|
ION Vital - Viability Assay Kit
Calcein AM is a membrane-permeant, non-fluorescent form of calcein that enters cells passively.
Once inside the cytosol of cells, Calcein AM is converted to green-fluorescent Calcein by
ubiquitous esterases in viable cells, resulting in uniform cytosolic fluorescence (Ex/Em 495
nm/515 nm). Calcein, a polyanionic dye, is membrane-impermeant and is well retained within
the cytosol of most healthy cells with intact cell membranes.
Ethidium homodimer I (EthD-I) is a membrane-impermeable, high-affinity, nucleic acid stain that
is excluded by viable cells with intact cell membranes. When membrane integrity is
compromised, a hallmark of dead or dying cells, EthD-I enters the cell and binds DNA, which
results in a >30-fold enhancement in bright red, nuclear fluorescence (Ex/Em 528 nm/617 nm).
ION Vital—Viability can be used to quantify live and dead cells within eukaryotic cell suspensions
or adherent cultures, 3D cultures, organoids, and some non-fixed tissues. Cannot be used for
yeast or bacteria.
When following our protocol, ION Vital—Viability kit provides enough reagents to make 150 mL of
working solution, enough for fifteen 96-well plates or 1,500 flow cytometry samples. The actual
number of assays will vary according to optimal dye concentrations for your application.
ION VITAL - Viability Kit
Kit Contents
|
Name
|
Volume
|
Containers
|
|
4mM Calcein AM in DMSO
|
100 µL
|
1
|
|
2mM Ethidium homodimer I (EthD-I) in DMSO/H2O
|
300 µL
|
1
|
Calcein Am
|
CAS number:
|
148504-34-1
|
Excitation:*
|
494 ± 3 nm
|
|
Emission:*
|
517 ± 3 nm
|
|
MW:
|
995 g/mol
|
|
Solubility:
|
DMSO
|
|
Purity (minimum):
|
≥ 95%
|
|
Storage:
|
-20o C, protect from light and moisture
|
References
1. Papadopoulos, N. G., Dedoussis, G. V., Spanakos, G., Gritzapis, A. D., Baxevanis, C. N., Papamichail, M. An improved fluorescence assay for the determination of lymphocyte-mediated cytotoxicity using flow cytometry. Journal of immunological methods. 1994. 177(1-2), 101–111.
2. A.W. Hayes, Ed., Principles and Methods of Toxicology, 6th Edition, Raven Press (1994) pp. 1231–
1258.
|
|
|
 |
|
 |
|