The use of high-performance liquid chromatography for the determination of size and molecular weight of proteins: a caution and a list of membrane proteins suitable as standards

• Published in: Analytical biochemistry Vol. 154; no. 2; p. 525
• Main Authors: Le Maire, M, Aggerbeck, L P, Monteilhet, C, Andersen, J P, Møller, J V
• Format: Journal Article
• Language: English
• Published: United States 01-05-1986
• Subjects: Calibration; Chemical Phenomena; Chemistry, Physical; Chromatography, High Pressure Liquid; Detergents; Membrane Proteins; Molecular Weight; Proteins - analysis; Reference Standards; Solubility; Ultracentrifugation
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(86)90025-4

We propose a list of 15 water-soluble globular proteins and 13 forms of detergent-soluble membrane proteins of known Stokes radii for the calibration of high-performance liquid chromatography columns. It is shown that it is advisable to use different sets of standards for these two types of proteins as the detergent-solubilized membrane proteins may behave differently, being excluded or retarded, depending upon the gel support. A smooth, although nonlinear, relationship between Stokes radii and erf-1(1--KD) is observed while a large scatter of points exists if the calibration is expressed as the molecular weight as a function of KD.