Unit

UNIT 5.5 Determination of Molecular Size by Size-Exclusion Chromatography (Gel Filtration)

  1. G. Brent Irvine

Published Online: 1 MAY 2001

DOI: 10.1002/0471143030.cb0505s06

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Irvine, G. B. 2001. Determination of Molecular Size by Size-Exclusion Chromatography (Gel Filtration). Current Protocols in Cell Biology. 6:5.5:5.5.1–5.5.16.

Author Information

  1. Queen's University Belfast, Medical Biology Centre, Belfast, U.K.

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: APR 2000

Abstract

Size-exclusion or gel filtration chromatography is one of the most popular methods for determining the sizes of proteins. Proteins in solution, or other macromolecules, are applied to a column with a defined support medium. The behavior of the protein depends on its size and that of the pores in the medium. If the protein is small relative to the pore size, it will partition into the medium and emerge from the column after larger proteins. Besides a protein's size, this technique can also be used for protein purification, analysis of purity, and study of interactions between proteins. In this unit protocols are provided for size-exclusion high-performance liquid chromatography (SE-HPLC) and for conventional gel filtration, including calibration of columns (in terms of the Stokes radius) using protein standards.