Nuclear Magnetic Resonance (NMR)
How To Get Started on an NMR Project
- Expression System/Cell types
- BL21(DE3)
- HMS174(DE3)
- We typically use pET expression vectors
- Appropriate Buffers
- Tris
- Hepes
- Phosphate
**if you have questions about a buffer, just give us a call** - 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)
- Things to buy:
- Sample Tubes:
- Labeled Isotopes:
- 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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