CyQUANT™ Direct Cell Proliferation Assay
CyQUANT™ Direct Cell Proliferation Assay
Invitrogen™

CyQUANT™ Direct Cell Proliferation Assay

Experience a more convenient workflow for your high-throughput cytotoxicity and proliferation screening projects with the CyQUANT™ Direct Cell Proliferation Assay–noRead more
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Catalog NumberQuantity
C350121 Kit(s)
C350111 Kit(s)
Catalog number C35012
Price (USD)
3,575.00
Each
Add to cart
Quantity:
1 Kit(s)
Price (USD)
3,575.00
Each
Add to cart
Experience a more convenient workflow for your high-throughput cytotoxicity and proliferation screening projects with the CyQUANT™ Direct Cell Proliferation Assay–no washes, cell lysis, temperature equilibrations, or radioactivity required. DNA-based assays have been shown to be among the most sensitive cell health indicators and data concordance with metabolically based assays is excellent. The CyQUANT™ Direct assay is a highly sensitive and robust method to assess cell growth, viability, or compound toxicity in a range of applications, from high throughput screening to bioproduction.

Features of the CyQUANT™ Direct Cell Proliferation Assay include:

• More convenient workflow for high-throughput screening–no washes, cell lysis, temperature equilibrations, or radioactivity required
• Rapid protocol, high sensitivity and dynamic range
• Measures independent of metabolic state of cells on standard fluorescent plate readers
• Multiplex assays using a spectrally distinct fluorescent or a luminescent readout

Rapid and convenient, with a large dynamic range
The CyQUANT™ Direct assay is a fluorescence-based proliferation and cytotoxicity assay for microplate readers with a linear detection range from less than 100 to 20,000 cells per well in most cell types. The no-wash, homogenous format and fast add-mix-read protocol makes the CyQUANT™ Direct assay ideal for HTS applications. The assay can be completed in one hour, with no washes, no cell lysis, or temperature equilibrations required. The signal is stable for several hours, affording additional work flow convenience.

How it works
The CyQUANT™ Direct assay is based on a cell-permeant DNA-binding dye in combination with a masking dye reagent. Cellular DNA is highly regulated and estimates of cell numbers based on DNA are very accurate. The masking dye blocks staining of dead cells or cells with compromised cell membranes resulting in only healthy cells being stained. The CyQUANT™ Direct assay, therefore, measures proliferation as well as membrane integrity, another measure of cell health. Users of metabolically based cell health assays can readily compare historical data with results from the CyQUANT™ Direct assay, as concordance assays based on metabolism or energy (ATP) has been shown to be excellent. Furthermore, it is possible to multiplex with spectrally distinct fluorescent probes or luminescence assay kits because cells are not lysed. Furthermore, the fluorescent signal allows the user to switch between HTS and HCS plate readers if multiplex assays require different platforms.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Cell TypeMammalian Cells
ColorGreen
Culture EnvironmentSuspension Cell Culture, Adherent Cell Culture
Detection MethodFluorescence
For Use With (Application)Proliferation Assay
For Use With (Equipment)Microplate Reader, Spectrophotometer, Microscope, HTS Reader
Product TypeCell Proliferation Assay
Dye TypeOther Label(s) or Dye(s)
EmissionVisible
Format96-well plate, 1536-well plate, 384-well plate
Incubation Time60 min
Product LineCyQUANT™, Molecular Probes™
Quantity1 Kit(s)
Shipping ConditionRoom Temperature
Unit SizeEach
Contents & Storage
  • 5 mL CyQUANT™ Direct nucleic acid stain (component A)
  • 25 ml CyQUANT™ Direct background suppressor I (component B)
  • Store both at 2°C to 25°C. Desiccate and protect from light.

Frequently asked questions (FAQs)

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.