Design of small volume HX and triple-resonance probes for improved limits of detection in protein NMR experiments

J Magn Reson. 2003 Sep;164(1):128-35. doi: 10.1016/s1090-7807(03)00184-8.


Three- and four-frequency nuclear magnetic-resonance probes have been designed for the study of small amounts of protein. Both "HX" (1H, X, and 2H channels) and "triple-resonance" (1H, 15N, 13C, and 2H) probes were implemented using a single transmit/receive coil and multiple-frequency impedance matching circuits. The coil used was a six-turn solenoid with an observe volume of 15 microl. A variable pitch design was used to improve the B1 homogeneity of the coil. Two-dimensional HSQC spectra of approximately 1mM single labeled 15N- and double labeled 15N/13C-proteins were acquired in experimental times of approximately 2h. Triple-resonance capability of the small-volume triple-resonance probe was demonstrated by acquiring three-dimensional HNCO spectra from the same protein samples. In addition to enabling very small quantities of protein to be used, the extremely short pulse widths (1H = 4, 15N = 4, and 13C = 2 micros) of this particular design result in low power decoupling and wide-bandwidth coverage, an important factor for the ever-higher operating frequencies used for protein NMR studies.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.
  • Validation Study

MeSH terms

  • Carbon Isotopes
  • Equipment Design
  • Lucanthone / analogs & derivatives*
  • Lucanthone / chemistry
  • Magnetics / instrumentation*
  • Microchemistry / instrumentation*
  • Microchemistry / methods
  • Nitrogen Isotopes
  • Nuclear Magnetic Resonance, Biomolecular / instrumentation*
  • Nuclear Magnetic Resonance, Biomolecular / methods
  • Proteins / chemistry*
  • Protons
  • Quality Control
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Transducers*
  • Ubiquitin / chemistry


  • Carbon Isotopes
  • Nitrogen Isotopes
  • Proteins
  • Protons
  • Ubiquitin
  • IA 3
  • Lucanthone