PrestoBlue™ Cell Viability Reagent is a ready‐to‐use reagent for rapidly evaluating the viability and proliferation of a wide range of cell types. PrestoBlue™ reagent is quickly reduced by metabolically active cells, providing a quantitative measure of viability and cytotoxicity in as little as 10 minutes. Protect reagent from long term exposure to light and avoid contamination, as this can reduce sensitivity.

Quick Reference Protocol

PrestoBlue™ Cell Viability Reagent

  1. PrestoBlue™ Cell Viability Reagent is supplied as a 10X solution. Add PrestoBlue™ Reagent  directly to cells in culture medium.  See below for example volumes:


    Format Volume of cells + media Volume of PrestoBlue™ Reagent
    Cuvette900 μL100 µL
    96-well plate90 µL10 µL
    384-well plate36 µL4 µL

    Note: Correct for background fluorescence by including control wells containing only cell culture  media (no cells) on each plate. 

  2. Incubate ≥10 minutes at 37º C. Longer incubation times will increase sensitivity of detection. As  this is a live cell assay, readings may be taken at multiple time points to determine optimal  performance in your lab.


    Format Recommended Incubation Time
    Bottom-read fluorescence10 minutes – 2 hours
    Top-read fluorescence30 minutes – 2 hours
    Absorbance20 minutes – 2 hours
    Room temperature incubation10 minutes – 2 hours
    Low cell number (< 5,000 cells/100 µL)20 minutes – 24 hours
  3. Read fluorescence or absorbance. Fluorescence is more sensitive than absorbance and is the  preferred detection method.

    Format Excitation Emission
    General540–570 nm 580–610 nm
    560 nm (10 nm bandwidth)590 nm (10 nm bandwidth)
    Fluorescence (Filter)535 nm (25 nm bandwidth)615 nm (10 nm bandwidth)
    Absorbance570 nm600 nm (reference wavelength for
  4. Calculate and plot the results. Higher fluorescence or absorbance values correlate to greater total  metabolic activity.


    Format Instructions
    • a. Optional: Average the fluorescence values of the no-cell control wells and
      subtract from the fluorescence value of each experimental well.
    • b. Plot fluorescence vs. experimental condition (cell number, compound
    • a. Normalize the 570 nm values to the 600 nm values for the experimental wells.
    • b. Plot the normalized 570 nm absorbance values vs. experimental condition (cell
      number, compound concentration).
MAN0003232       11–Oct–2010