Gen5 version required: 1.0 or higher
Basis for the Assay:
The internal environment of proliferating cells is more reduced than that of non-proliferating cells. Compounds such as tetrazolium salts and alamarBlue™, which can be reduced by cellular metabolic intermediates, can be used to monitor cellular proliferation. The reduction of AlamarBlue results in a measurable color shift. The wavelengths for maximal absorbance are 570 nm and 600 nm for the reduced and oxidized forms of alamarBlue respectively. Because the wavelengths overlap it is necessary to measure the absorbance at both wavelengths.
Protocol name: AlamarBlue-A.prt (Absorbance Readers)
The protocol calls for absorbance measurements at 570 nm and at 600 nm.
Plate configuration provides for samples to be run in duplicate. In addition, two blank wells (Media only) and two control-wells (media plus alamarBlue) are defined on the plate. In regards to data reduction, a transformation that calculates reduction percentage using the molar extinction coefficients of reduced and oxidized alamarBlue at 570 nm and 600 nm has been created.
The report is configured to provide the (1) Blanked absorbance at 570 nm and 600 nmf and (2) Percent reduction calculation.