Table 2 Tetrazolium and alamar blue assay conditions.

From: Variation in pentose phosphate pathway-associated metabolism dictates cytotoxicity outcomes determined by tetrazolium reduction assays

Method

Proportion

dye:medium

Incubation duration

Quantitation method

WST1

1:9

2 h

Absorbance1

Measure: 450 nm

Reference: 650 nm

MTS

1:4

2 h

Absorbance1

Measure: 490 nm

MTT

1:4

4 h *

Absorbance1

Measure: 570 nm

Reference: 600 nm

Alamar Blue

1:9

3 h

Fluorescence2

Excitation: 560

Emission: 590

  1. *Insoluble MTT formazan was resolubilized in neat DMSO
  2. 1SpectraMax Plus 384 and SoftPro Max data acquisition software (Molecular Devices; Le Jolla, California)
  3. 2Varioskan LUX and SkanIt v. 4.1 data acquisition software (Thermo Fisher Scientific, Inc.).
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