I am trying to find the best method to determine the effects of adding an inhibitor on the function of one protein ( transmembrane) in cell culture.
Thank you!
Soner is exactly right - knocking down the protein with siRNA is a standard procedure (see examples at the bottom here: https://altogen.com/) for looking at the effects of protein inhibition. Adding protein-blocking aptamers is another method, but siRNA-based silencing is a well-researched technique that has been crucial to many studies.
Hi Monica,
I'd think that knocking down the gene of interest by siRNA is the most promissing approach.
Best regards
Soner is exactly right - knocking down the protein with siRNA is a standard procedure (see examples at the bottom here: https://altogen.com/) for looking at the effects of protein inhibition. Adding protein-blocking aptamers is another method, but siRNA-based silencing is a well-researched technique that has been crucial to many studies.
Dear Monica
1. Adding inhibitor (e.g. siRNA) in cell culture
2. Real-time PCR /western blot
Best
Hi,
If your protein has special function, you can use indirect assay for evaluation of inhibitor efficacy. For example, if your protein is a trans-membrane enzyme , Km and Vmax if enzyme after and before of inhibitor challenge maybe good index for your purpose.
Thank you all for your very helpful replies. I think I will try knocking down the protein with siRNA and see how it goes.
Thank you!
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