Alamar Blue Assay

Alamar blue is based on the chemical of resazurin

Invitrogen: alamar blue cell viability reagent

10x, ready to use solution
25mL for 2500 assays

Preparing Cells:
  • adherent mammalian cells- plate the cells and allow cells to adhere and grow approximately 4- 24 hours at 37℃ and 5% CO2 before proceeding with the assay
  • suspension mammalian cells-plate the cells and use cells immediately for the assay, or alternatively allow the cells to grow for up to 24 hours at 37℃ and 5% CO2 before proceeding with the assay.
  • Bacterial cells:

#treat cells with the testing compound for 24-72 hours prior to performing the alamar blue assay
1 add 10th volume of alamar blue to cells in culture medium as described below
Cuvette:1mL of cell suspension + 100µL alamar blue
96-well:100µL of cell suspension + 100µL alamar blue
384-well:40µL of cell suspension + 100µL alamar blue
1 to 4 hours incubation at 37℃ in incubator (protect from direct light)

  1. Fluorescence: red fluorescence Ex.540-570nm(570), Em.580-610nm(585nm)
  2. Absorbance: monitor the absorbance at 570nm and 600nm
optional:reaction stop by 3% SDS in PBS, pH 7.4
1. fetal bovine serum and bovine serum albumin cause some quenching of alamar blue fluorescence. recommended using the same serum concentration in controls to account for this quencing. Other media components such as phenol red do not interfere with the assay.

AbD serotec

determination of optimal cell density and incubation time for A549 cells


CellTiter-Blue Cell Viability Assay

S. Al-Nasiry et al., the use of alamar blue assay for quantitative analysis of viability, migration and invasion of choriocarcinoma cells, human reproduction, 2007, 22(5), 1304-1309

Sherry L. Voytik-Harbin, et al., Application and evaluation of the alamar blue assay for cell growth and survival of fibroblasts, In vitro cellular and developmental biology - animal, 34(3), 239-246