Sandwich-ELISE NMR: reducing the sample volume of NMR samples.
We present Sandwich-ELISE, a concatenated version of our previously proposed Experimental LIquid SEaling (ELISE) protocol, in which an aqueous sample is effectively sealed by the addition of a small layer of mineral oil, or, alternatively, a chloroform sample was sealed by a water layer. With Sandwich-ELISE, a triple layered geometry composed of deuterated chloroform/aqueous buffer/mineral oil can be used to limit the sample to the active coil volume, effectively replacing the popular Shigemi tubes. Importantly, this procedure is readily applicable to smaller diameter tubes, for which no Shigemi tubes are available. We further present spectra of a 1 microl protein sample sandwiched between the chloroform and Nujol phases in a 1mm tube, demonstrating thereby that the volume of the aqueous phase of interest can be reduced even further.
Wieruszeski
Jean-Michel
JM
CNRS, Université de Lille 1, UMR 8576, USTL, Avenue de Mandeleiev, FR-59655, Villeneuve d'Ascq Cedex, France.
Fritzinger
Bernd
B
Hanoulle
Xavier
X
Martins
José C
JC
Lippens
Guy
G
D002725:Chloroform / Q000737:chemistry; D021983:Cyclophilins / Q000737:chemistry*; D008899:Mineral Oil / Q000737:chemistry; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D012815:Signal Processing, Computer-Assisted; D012996:Solutions / Q000737:chemistry*; D013395:Sucrose / Q000737:chemistry
D012996:Solutions; D013395:Sucrose; D002725:Chloroform; D008899:Mineral Oil; D021983:Cyclophilins