AquaBluerTM is a proprietary redox indicator. Viable cells turn AquaBluer from its oxidized form (nonfluorescent, blue) to the reduced form (fluorescent, red). The fluorescence intensity of AquaBluer at 540ex/590em is proportional to the number of viable cells in the sample. It can be used to assess cell viability, cell proliferation, and cytotoxicity. It is nontoxic, simple to use, sensitive, reproducible, and has a broad assay range. Most importantly, it reduces your assay cost 3 folds over Alamar Blue and 26 folds over MTT.
Perform viability assay with a fluorescence plate reader or absorbance plate reader.
Just add AquaBluer to the culture, incubate and read. No PBS, no SDS and no DMSO.
Not a terminal assay. AquaBluer-treated cells may still be used in other analyses.
More saving means more research to you.
Comparison of Alamar Blue, AquaBluer and MTT Assays
AquaBluer has a wider dynamic assay range than Alamar Blue
1. Set up a cytotoxicity assay
Seed the cells at 6000-8000 cells/100 ul/well in 96-well culture plates and let the cells grow overnight. Add serially diluted test compounds to each well. Include 4 “No-cells” and 4 “No-drug” wells as 0% and 100% viability controls. Return the plate to the CO2 incubator for 48-72 hrs.
2. Start AquaBluer assay
Add 100 ul AquaBluer solution (15 ml in the kit for 15,000 assays) to 10 ml culture medium in a dispensing reservoir. Pipette up and down to mix well. Replace the culture medium in each well with the 100X diluted AquaBluer medium. Alternatively, directly add 1 ul of undiluted AquaBluer solution to each 100 ul well, especially for viability assay of suspension cultures. Return the plate to the CO2 incubator for 4 hrs.
3. Raw data acquisition
Remove the plate from the incubator. Place the plate in a fluorescence plate reader and read the fluorescence intensity at 540ex/590em.
4. Viability and IC50 calculation
Subtract the average of fluorescence value (RFU) of the No-cell control (background) from all other RFU values. Convert the test RFU values to % viability using the formula:
% Viability = (RFUtest / RFUveh) x 100,
where RFUveh is the average RFU of the No-drug wells. Enter the % viability values and corresponding log test compound concentrations into a non-linear regression program such as Prism and use the Four Parameter Model to obtain the IC50 values and dose-response curve of the test compounds.
Product Name: AquaBluerTM Kit
Product Number: 6001, 6015
Application: Assessing cell viability, cell proliferation, and cytotoxicity.
Size: The kit is sufficient for performing
Kit Contents: The AquaBluer Kit includes the following items
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