Gel filtration chromatography (size exclusion chromatography) of proteins

Krisna C Duong-Ly; Sandra B Gabelli

doi:10.1016/B978-0-12-420119-4.00009-4

Gel filtration chromatography (size exclusion chromatography) of proteins

Methods Enzymol. 2014:541:105-14. doi: 10.1016/B978-0-12-420119-4.00009-4.

Authors

Krisna C Duong-Ly¹, Sandra B Gabelli²

Affiliations

¹ Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
² Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Electronic address: gabelli@jhmi.edu.

PMID: 24674066
DOI: 10.1016/B978-0-12-420119-4.00009-4

Abstract

The protocol described here allows the student to construct a standard curve for a gel filtration column with a separation range of 5-250 kD. The size (hydrodynamic radius) of a protein species stable in a buffer containing Tris-HCl, NaCl, and DTT is determined using this column. Modifications may be made to the buffer to accommodate the protein of interest and the separation range of the column.

Keywords: Column volume; Gel filtration chromatography; Gel filtration column; Size exclusion chromatography; Void volume.

MeSH terms

Buffers
Chromatography, Gel / instrumentation
Chromatography, Gel / methods*
Proteins / isolation & purification*
Tromethamine / chemistry

Substances

Buffers
Proteins
Tromethamine