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Order ProtocolMSDSFAQAquaGenomicAquaBluer

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AquaBluer is a proprietary redox indicator for cell viability and cytotoxicity assays. 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.




AquaBluer is a colorimetric and fluorescent redox indicator. Cell viability and cytotoxicity assays may be performed with a fluorescence plate reader or absorbance plate reader.


Assay is performed in culture medium. Just add AquaBluer to the culture, incubate and read. No PBS, no SDS and no DMSO.
Nontoxic, sensitive, reproducible, and with a broad assay range

Not a terminal assay. AquaBluer-treated cells may still be used in other analyses.


Costs 3X less than Alamar Blue assay and 26X less than MTT assay. More saving means more research to you.


Comparison of Alamar Blue, AquaBluer and MTT Assays

   Alamar Blue  AquaBluer  MTT
 Detection mechanism  Redox indicator  Redox indicator  Redox indicator
 Protocol 1. Replace culture medium with PBS-based Alamar Blue.

2. Incubate for 4 hrs
(Your cells are not happy to be fasting in PBS for 4 hrs and their metabolism may be altered artificially!)

3. Read.
1. Replace culture medium with medium-diluted AquaBluer or just add undiluted AquaBluer to the culture.

2. Incubate for 4 hrs (Your cells remain in their accustomed culture medium during the assay.).

3. Read
1. Replace culture medium with medium-diluted MTT.

2. Incubate for 4 hrs.

3. Add SDS-HCl and pipet to lyse the cells. 

4. Incubate for 4 hrs to solubilize the formazan dye.

5. Pipet to mix thoroughly and read.

 Price per kit  $400 /100 ml  $199 /15 ml  $340 /10 ml
 # of assays per kit  10,000  15,000  1,000
 Assay range  600-75,000 cells  600-150,000 cells  2,000-250,000 cells

AB assay

AquaBluer has a wider dynamic assay range than Alamar Blue

Brief Protocol

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, convenient for viability assay of fresh 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.

Order Information

Product Name:    AquaBluerTM Kit 

Product Number:   6001, 6015

Application:    Assessing cell viability, cell proliferation, and cytotoxicity. For in vitro research use only.

Size:    The kit is sufficient for the preparation of

  • 6001: 1,000 assays in 96-well format using 1 ul AquaBluer in 100 ul medium
  • 6015: 15,000 assays

Kit Contents:   The AquaBluer Kit includes the following items

  • 6001: 1 ml AquaBluer solution, User Manual
  • 6015: 15 ml AquaBluer solution, User Manual


  • 6001: $10 each
  • 6015: $199 each
Ordering:    To order, please click the "BUY" button below. 



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