I use ELISAs for quantification of extracellular proteins in cell-conditioned media. I am interested in quantifying uncoupling protein 1 (UCP1) which is an intracellular, mitochondrial membrane-associated protein. I like to use TRI/Trizol reagent for RNA, DNA, protein extraction (I am old school--plus it works well and it's cheap). Does anyone know if extracting protein with the TRI method would yield proteins that can be used in ELISAs? The proteins will be solubilized and stored in 1% SDS, according to the protocol.
Yes, you can use proteins extracted with the TRI/Trizol method for ELISAs, but there are key considerations to remember. SDS, even at 1%, can disrupt antibody binding in ELISA assays. Here are some tips to improve compatibility:
By following these steps, you can successfully prepare your TRI/Trizol-extracted proteins for ELISA quantification. This approach should help optimize your results.
Good luck!
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