I’m looking for a fluorescence-based assay for quantification of low molecular thiols in the nM range in a cell sample. I find numerous commercially available kits and since they all claim to be proprietary without giving the name of the probe it’s hard to compare. Which assay for quantification of thiols in a cell extract do you find to be the most sensitive and stable?
Hi, I would use a CPM (7-Diethylamino-3-(4'-Maleimidylphenyl)-4-Methylcoumarin) based assay. The probe has a low fluorescent active, but the thiol-probe conjugate is very active (384/470nm). I've only used it for a HPLC based assay, but I'm sure you would be able to use it for a plate based assay (to determine the total thiol count) as well.
I was also wondering about such method. I'd like to thank Dr. Lund for the question and Dr.Lamprecht for the answer.
I used Ellmans reagent which is colorimetric. The protocol is on the internet.
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