I have some brain and adipose tissue from rats that I would like to run qPCR and ELISA on, using this TRIzol isolation protocol first (https://tools.thermofisher.com/content/sfs/manuals/trizol_reagent.pdf).
The protocol says you can run western blot on the isolated protein but I can't seem to find anything on whether you could run ELISA instead of WB?
If anyone has experience of doing this or just some advice it would be very much appreciated!
TRIzol denatures proteins. Your ELISA capture and detecting antibodies may or may not work properly. I would initially run an ELISA trial treating the standards with and without TRIzol.
Hi Gordon,
Thanks for your advice. I also contact the supplier (ThermoFisher) and they said 'We would not recommend to use proteins isolated with Trizol for ELISA. ELISA is not usually performed with lysate samples like that coming from Trizol isolation, since not all proteins might have the native conformation needed.'
I think I'm just going to run ELISA first and if I have any tissue left over I will run qPCR.
Thanks,
Anna
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