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What if NMR Is Not For You?

I am a lifelong user of nuclear magnetic resonance (NMR) spectroscopy to study peptides and proteins. Having written a ton of papers, having trained hundreds of students to use NMR to study peptides and proteins, and having worked all sorts of job positions in my career, I feel pretty confident that NMR works really well for a wide range of proteins. Nevertheless, each time I announce to a new student that I use NMR to study proteins, I get one or two people who are interested in using NMR for their own protein research.

Unfortunately, NMR is really not for everyone. I never tell anybody that they cannot use NMR to study their proteins – everybody can. The problem that people experience, however, is that they cannot get good NMR spectra from their proteins. Lots of people have tried lots of things. They have tried everything – changing buffer conditions, changing the pH, changing the salt concentration, replacing temperature to test if they can get a spectrum. They have even tried different NMR setups, from benchtop to full size spectrometers. They may try to understand why they cannot get a spectrum. The problem is, they simply do not know.

Some people have even approached me at scientific meetings to ask me to explain this to their graduate students. People always ask me whether they are doing anything wrong with their protein samples. The answer is no, they are doing it all right. I always try to explain that NMR may not be for them, but they need to find out why for themselves.

I like to share that I have a small collection of NMR spectra from people who once told me they cannot get any NMR spectrum from their protein sample. They all find out why on their own. These are the moments that I really treasure. This is a collection that I always share with my students at the end of my course on protein NMR.

This is what I advise to anyone who is trying to do NMR experiments on their own proteins.

1. Find out the conditions that are best for your protein. For my simple examples, I usually find out the conditions that produce the best NMR spectra for my samples. Find out what the best concentration is for your sample that gives you the best spectrum. Usually, it is around 1 mM but I have also seen spectra from 0.

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