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.
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 or online calculator such as BioDataFit at http://www.changbioscience.com/stat/ec50.html
to obtain the IC50 values of the test compounds.
Ordering
Information
Product
Name: AquaBluerTM
Kit
Product
Number: 6001, 6015
Application:
Assessing
cell viability, cell proliferation, and cytotoxicity.
Size:
The kit is sufficient for performing
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,
Instruction Manual
6015: 15 ml AquaBluer solution,
Instruction Manual
Price:
6001: $10 each
6015: $199 each
Ordering:
To order, please
click the "BUY" button below.
