Nuclear Magnetic Resonance (NMR)

How To Get Started on an NMR Project

1. Expression System/Cell types
• BL21(DE3)
• HMS174(DE3)
• We typically use pET expression vectors


2. Appropriate Buffers
• Tris
• Hepes
• Phosphate
  **if you have questions about a buffer, just give us a call**


3. Sample Conditions
• Protein Concentration: We recommend a protein concentation of between 0.2mM - 2mM (less than 0.2mM is possible for some experiments if necessary)
• Sample Volume: Samples need to be 600ul if using a standard NMR tube, or no less than 300ul if using a Shigemi tube
• D2O: The final NMR sample must contain between 5% and 10% D2O
• Salt Concentrations: Try to keep this LOW (<100mM total ionic strength)


4. Things to buy:
• Sample Tubes:
• Shigemi
• Wilmad
• Labeled Isotopes:
• CIL for 13C-glucose
• CIL for 15N-ammomium sulfate
• CIL for D20

1H-15N HSQC of TRTK-bound S100B

The above image illustrates typical preliminary data collected at the start of a protein NMR project. The above spectrum takes approximately 15 minutes to collect and tells much about the feasibility of a given NMR project. NMR structural work is carried out on proteins grown with nutrients labeled with both 15N and 13C.

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