Alamar blue protocol - (Nov/30/2006 )
Has any one done the cell viability /cell toxicity assay using alamar blue. The protocol with the reagents is not very clear and cites references. The references in turn cite the protocol . So one ends up going round and round without any clear conclusion.
If any one has actually used this particular reagent for the viability/toxicity assay, Please help me out. I'd be much obliged.
thanks and take care
If any one has actually used this particular reagent for the viability/toxicity assay, Please help me out. I'd be much obliged.
thanks and take care
I've used alamar blue in the past. It's really very simple. Just dilute into your cell growth media and place on your cells. You will have to determine the length of time for development though. Watch for color change from blue to pink. Don't let the color get really pink or you will have saturated the assay. I like using a fluorescent reader for the assay, but you can also use colorimetric reading at two wavelengths (I forget the exact formula for this). Make sure to have a negative control (no cells) for background subtraction.
From my understanding Alamarblue is similar or the same as Resazurin. If so, I've used it before and it is very easy to use. I use a 96-well TC plate with 100 uL of media in each well. When the desired amount of time has passed for your assay all you do is add 10 uL/well Resazurin (1mg/mL in PBS) to each well. Then, incubate for 2-3 hours at 37ºC. The excitation and emission wavelengths are 485 and 530 nM.