MTT cell proliferation plate assays require cellular metabolism to modify the reagent and thus, only live cells will be counted. Is this true for CyQUANT Direct Cell Proliferation Assay, too?
This is true for MTT (as well as XTT, AlamarBlue Cell Viability Reagent, and PrestoBlue Cell Viability Reagent). CyQUANT Direct will also only count live cells, but for a different reason. The dye in it is a green-fluorescent nucleic acid stain, which will bind to DNA in all cells, live or dead, without the need for cellular metabolism. However, there is a non-cell-permeable quenching reagent in the kit which will both quench extracellular fluorescence (and thus this is a no-wash assay) AND will quench the fluorescence in dead cells (but not live cells).
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
MTT cell proliferation assays require cellular metabolism to turn over the reagent, and thus only live cells will be counted. Is this true for CyQUANT Direct as well?
This is true for MTT (as well as XTT, AlamarBlue Cell Viability Reagent, and PrestoBlue Cell Viability Reagent). CyQUANT Direct will also only count live cells, but for a different reason. The dye is a green-fluorescent nucleic acid stain, which will bind to DNA in all cells, live or dead, without the need for cellular metabolism. However, there is a cell impermeant quenching reagent in the kit which will quench both extracellular fluorescence (and thus this is a no-wash assay) and the fluorescence in dead cells (but not live cells).
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
What volumes should I use to make a 10X stock solution for the CyQUANT Direct Cell Proliferation Assay (Cat. No. C35011, C35012)?
To make a 10X stock solution for the CyQUANT Direct Cell Proliferation Assay (Cat. No. C35011, C35012), we suggest using the following volumes:
1.1 mL PBS
25 µL CyQUANT Direct nucleic acid stain (Component A)
125 µL CyQUANT Direct background suppressor I (Component B)
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Are the components of the CyQUANT Direct Red Cell Proliferation Assay and CyQuant Direct Cell Proliferation Assay kits interchangeable?
No. The components of the CyQUANT Direct Red Cell Proliferation Assay and CyQuant Direct Cell Proliferation Assay kits are not interchangeable.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
What is the difference between the CyQUANT Direct Red Cell Proliferation Assay (Cat. Nos. C35013, C35014) and the green fluorescent CyQuant Direct Cell Proliferation Assay (Cat Nos. C35011, C35012)?
The two kits utilize different dyes and background suppressors, but they function in the same way and provide similar results as can be seen from Figures 1 and 2 in the CyQUANT Direct Red Cell Proliferation Assay product sheet (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0018644_CyQUANT_direct_red_assay_PI.pdf). Dyes from both kits are nucleic acid stains and the background suppressors are cell impermeant reagents that quench extracellular fluorescence. They are usable within the same range (100 to 20,000 cells/well in a 96-well format) and in the same workflow.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.