Usually, a 96 well plate is recommended for WST-1 assay, but its difficult sometimes. So, after performing this assay in a 12 or 24-well plate, is it possible to measure absorbance using a 96 well plate?
If u have a reader for those plates its possible (I know readers with 24well plate possibility). But Its money loss and extra work, with less replicates possibility
Yes, Asadur,, it is. We do it in our lab at a regular basis. We use the 24 well format because our cells come from an experimental set up where the cells are attached to round coverslips that fit into the 24-well plate. You only need a plate reader that will read the 24-well format. We use a final volume of 300 uL (270 uL medium+30 uL WST-1 reagent). As pointed above, it's costly, but ti fits some purposes very well.
Yes, if you have a PTP reader for this plates. Only add 10% of WST-1 to you media!
If you do not have a 24 well plate reader, you can try the following method which we use in our lab; perform the experiment in 24 well or a 12 well plate, once your assay is complete trypsinize the cells and count them. Approximately add 20000 cells per well in a 96 well plate and add fresh media to make the total volume to 200 micro liters. Now add the WST-1 reagent according to the manufacturer's protocol and take a reading
yes it's possible...if you don't have an appropriate plate reader you can divide the medium with WST in a 96 well plate and then measure absorbance, but remeber at the end to average the obtained measures.
We regularly do this. We add the WST-1 to our cells in 200 µL in a 24 well plat (just 8-10µL depending on how fast your cells are) and after developing we remove 100µL, transfer it to a 96 well plate and measure it. We usually do this for adherent cells that don't like to be trypsinized.
Titrating the WST-1 for your cells is also worth a try we never need the amount that is given in the manual.
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