Calibration of gel-permeation columns in the high-molecular-mass range: fixed human thrombocytes for the estimation of interstitial volume and the haemocyanin of the Vineyard snail Helix pomatia as a molecular mass calibration substance

Abstract

Human thrombocytes, fixed with formaldehyde, are shown to be a universal indicator of the interstitial volume m gel-permeation chromatography (size-exclusion chromatography) for gels of all molecular mass separation ranges. The fixed thrombocytes are simple to prepare and to handle. Furthermore, native or intramolecularly cross-linked haemocyanin of the Roman snail Helix pomatia is shown to be well suited as a molecular mass calibration substance. We propose 8.8 · 10 6 as its molecular mass for calibration. This haemocyanin can therefore be added to the list of globular protein molecular mass calibration substances in the range of very high molecular masses.