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PrestoBlue™ Cell Viability Reagent

PrestoBlue™ Cell Viability Reagent


PrestoBlue™ Cell Viability Reagent
  • Save time with an incubation step as short as 10 minutes
  • Obtain high quality results with large assay window and z’ values
  • Experience convenience with an add-and-read homogenous format
  • Monitor living cells and perform downstream functional assays with no cell lysis
       
 
Ordering Information

What is PrestoBlue™ Reagent?

PrestoBlue™ reagent is a resazurin-based solution that functions as a cell viability indicator by using the reducing power of living cells to quantitatively measure the proliferation of cells. The PrestoBlue™ reagent contains a cell-permeant compound that is blue in color and virtually nonfluorescent. When added to cells, the PrestoBlue™ reagent is modified by the reducing environment of the viable cell, turns red in color and becomes highly fluorescent. This change can be detected using fluorescence or absorbance measurements.

Assay Workflow Complete in 10 Minutes

A 96-well plate containing the cells and the compounds to be tested is prepared.The PrestoBlue™ reagent is added directly to cells and incubated at 37°C for 10 minutes.The plate is then transferred to a fluorescence reader to measure signal.Results are evaluated by plotting signal versus compound concentration (Figure 1).In addition to this protocol format, the signal can alternatively be detected by absorbance or the assay can be run at room temperature.(View product protocol for full details on these assay variations.)

PrestoBlue-reagent

Figure 1. PrestoBlue™ cell viability reagent protocol.


Reliable, High Quality Results

PrestoBlue-reagent
 

Data obtained using PrestoBlue™ reagent shows excellent linearity (R2 value of 0.9948) for cells plated at densities between 100 and 100,000 cells/well.The signal for 100 wells was greater than that from zero cells +/- 3 standard deviations, and the Z’ values were above 0.5 for cell densities of 391 cells/well or greater (Figure 2).
Figure 2. Excellent data obtained in just 10 minutes.Jurkat cells were plated in cell culture medium in a 384-well plate in quadruplicate starting at 100,000 cells/well with a two-fold serial dilution.Cells were incubated with PrestoBlue™ reagent for 10 minutes prior to fluorescence measurements.


Sensitive Detection of Cell Viability
PrestoBlue-reagent
 

At 10 minutes of incubation, PrestoBlue™ reagent can detect as few as 98 cells/well, and the signal remains linear over the a broad dilution range.Following a 16-hour incubation with PrestoBlue™ reagent, as few as 12 cells/well exhibited a signal that was greater than that from zero cells +/- 3 standard deviations. Z’-factor values were above 0.5 for cell densities of 24 cells/well or greater (Figure 3).Note that at low cell densities, the signal-to-background ratio increases after 16 hours’ incubation.


Figure 3. Extremely sensitive detection.Jurkat cells were plated in cell culture medium in a 384-well plate in quadruplicate starting at 100,000 cells/well with a two-fold serial dilution.Fluorescence measurements were taken following 10 minutes of incubation and following 16 hours of incubation with PrestoBlue™ reagent.

PrestoBlue™ Reagent for Primary and Stem Cells

PrestoBlue™ Reagent is Compatible With Primary Cells

PrestoBlue-reagent

Cells remain viable so you can perform downstream functional assays with your cells after you have assessed their viability status.PrestoBlue™ reagent is the ideal option when using cells available in a limited supply such as primary cells (Figure 4).



Figure 4. PrestoBlue™ reagent enables analysis of primary cells. PrestoBlue™ reagent has been tested using human aortic smooth muscle cells. 

PrestoBlue™ Comparison to Other Resazurin-Based Viability Assays

PrestoBlue™ Reagent Viability Assays Are Significantly Faster Than Rezasurin-based Viability Assays, Without Any Loss of Sensitivity

PrestoBlue-reagent
PrestoBlue™ reagent only requires a 10-minute incubation period, whereas alamarBlue® and CellTiter-Blue® recommend a 1 to 4 hours incubation.Figure 5 demonstrates the difference in fluorescence signal obtained from PrestoBlue™ reagent and CellTiter-Blue® reagent following a 10-minute incubation.The dynamic range is significantly greater at the early time points for PrestoBlue™ reagent.


Figure 5. Achieve assay results with PrestoBlue™ reagent in a fraction of the time.CHO-K1 cells were plated in cell culture medium in a 384-well plate in quadruplicate starting at 10,000 cells/well with a two-fold serial dilution.Cells were incubated with PrestoBlue™ reagent, alamarBlue® reagent, or CellTiter-Blue® reagent for 10 minutes or 4 hours.Fluorescence measurements obtained with PrestoBlue™ reagent in 10 minutes take 4 hours with alamarBlue® and CellTiter-Blue® reagents.

PrestoBlue™ Reagent Compared to Tetrazolium Salt-Based Viability Assays

The PrestoBlue™ Reagent Has Many Advantages Over Tetrazolium Salts Like MTT.

  • Faster time-to-results and fewer protocol steps (Table 1)
  • Ability to maintain your cells in a living state for downstream assays
  • A safer reagent that minimizes toxic waste considerations
  • A ready-to-use aqueous solution

Shorter Assay Times

  MTT XTT PrestoBlue™
Minimum incubation time 1 小时 1 小时 10 分钟
Solubilization time 1 小时 N/A N/A
Total time 2 hrs 1 小时 10 分钟


Table 1.
Reduced time-to-results with PrestoBlue™ reagent.With PrestoBlue™ reagent, results can be obtained in a minimum of 10 minutes as compared to a minimum of 1 hour for XTT or 2 hours for MTT.

PrestoBlue™ Comparison to CellTiter® Reagent

Continuous Live-cell Monitoring and Assessment After Measuring Viability
PrestoBlue-reagent

 

Comparable results obtained with PrestoBlue™ reagent or CellTiter-Glo® reagent in 10 minutes.PrestoBlue™ reagent does not require cell lysis like the CellTiter-Glo® reagent does.This allows you to leave PrestoBlue™ reagent with the cell samples and read the signal up to 24 hours later.With PrestoBlue™ reagent, cells remain viable so you can perform downstream functional assays with your cells after you have assessed their viability status.If you are using cells that are in limited supply such as primary cells, PrestoBlue™ reagent is an excellent choice.

Figure 6. Comparable results obtained with PrestoBlue™ reagent or CellTiter-Glo® reagent in 10 minutes.U-2 OS cells were plated in a 384-well plate at 2,000 cells/well.Cells were then exposed to various concentrations of Etoposide, Doxorubicin, Chetomin, or Staurosporine for 72 hours.Subsequently, cells were incubated for 10 minutes with PrestoBlue™ reagent (left panel) or CellTiter-Glo® reagent (right panel) prior to fluorescence (PrestoBlue™ reagent) or luminescence (CellTiter-Glo® reagent) measurements.Comparable EC50 values were obtained for each compound.
 
 
仅用于研究用途。不得用于任何动物或人类的治疗或诊断。

CellTiter®-Glo and CellTiter®-Blue are registered trademarks of Promega Corporation.
alamarBlue® is a registered trademark of TREK Diagnostic Systems.
 
 
 
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