The AU-CD beam line on ASTRID2
In any CD experiment in general, and especially for a SRCD experiment, great care in the choice of buffers and salts should be taken. Chloride for example is one of the ions which should be avoided due to high absoprtion in the Far-UV/VUV. This means that for example Tris buffers which are pH regulated with HCl must be avoided, and NaCl salt should be substituted with NaF whenever possible. A few guidelines are listed below.
- Lowest wavelength data are obtained using short pathlength cells. We have cells with path lengths down to about 10mu.
- If possible, use pure deionized water as a solvent.
- Phosphate buffers are well suited to SRCD measurements, but concentrations should be kept below 20mM.
- NaCl salt should be avoided. If possible use NaF instead to obtain ionic strength.
For a full review of the optimum experimental conditions for SRCD we recommend reading chapter 4 in:
Miles, A.J. and Wallace, B.A. "Synchrotron Radiation Circular Dichroism Spectroscopy of Proteins and Applications in Structural and Functional Genomics". Chem. Soc. Reviews 35 (2006) 39-51.
A general review on how to do CD can be found in:
Kelly, S.M.; Jess, T.J.; Price, N.C. "How to study proteins by circular dichroism" Biochimica et Biophysica Acta 1751 (2005) 119 – 139.
For further information on and access to the AU-CD beamline please contact:
Last Modified 07 August 2014