I am doing time resolved FTIR between a protein and CBD. My protein is in HEPES buffer - but the spectrum only picks up the buffer and not my protein?

30 June 2023 1 4K Report

I am doing time resolved FTIR between a protein and CBD. My protein is at 66uM and the CBD at 1250uM - both in Hepes buffer (20mM hepes, 200mM NaCl pH 7). I tried a lower concentration of CBD in hepes (50uM) but FTIR did not pick up the CBD. Only with a really high concentration did I see a difference. However I am trying to test the interaction between CBD and my protein - but I do not get a protein spectrum - I only get a spectrum of the HEPES. I am not sure how to troubleshoot this experiment.

Paul Milham

It sounds like a sensitivity issue. Can you try increasing the protein concentration and developing a 'calibration'?

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