Experimental considerations fo the NMR Spectroscopy experiments

NMR spectroscopy is a multifaceted technique that enables you to address a great range of research problems in analytical chemistry, structural biology and biophysical chemistry. Below we outline just a few examples of projects that we have addressed in the past.
To meet all demands and improve the success rate of NMR spectroscopic projects, we routinely use the high-throughput capabilities of our crystallization platform to screen for optimal solution conditions using the method of
Differential scanning fluorimetry (DSF) used by structural genomics projects. Please contact us to discuss your specific needs and ideas!

Considerations for protein NMR experiments
The following points are guidelines that define a typical high-quality sample for protein NMR. Many applications of NMR spectroscopy work around these limitations. Please inquire for advice on your particular project.

Labeling with stable isotopes: Protein NMR spectroscopy often require stable isotope enrichment, e.g. 15N, 13C, and 2H. Expression in bacterial cultures grown on minimal media usually offers a cost-effective option.

For high-resolution tertiary protein structure determination by NMR spectroscopy, the molecular weight of the protein is typically less than 30 kDa. Larger proteins (up to 100 kDa in favorable cases) can be studied in projects that aim at identifying ligand binding sites or characterizing the physical state of the construct.

Depending on the size and solubility of the protein, for biological NMR spectroscopy a sample typically contains 1–10 mg/ml of purified protein in a suitable buffer.

For most in-depth studies by NMR spectroscopy, the protein needs to be stable for at least several days, and preferably for weeks or months. By identifying optimal solvent conditions, it is often possible to achieve a dramatic improvement in long-term stability, which leads to high quality protein NMR data.
For more details on NMR spectroscopy services please view our respective
services page.

Custom NMR spectroscopy services provided by SARomics Biostructures include:
- High-resolution 3D structure determination in solution
- Ligand binding assay and fragment screening
- Verification of target protein constructs
- pKa determination of titrating groups
- Validation of peptides, peptidomimetics, hormones, and other low-molecular-weight compounds