Preparation of membrane fractions from human neutrophil granules : A simple method

• Published in: Methods in cell science Vol. 19; no. 3; pp. 197 - 205
• Main Authors: VITA, F, BORELLI, V, SORANZO, M. R, MAGNARIN, M, BERTONCIN, P, ZABUCCHI, G
• Format: Journal Article
• Language: English
• Published: Norwell, MA Kluwer 1997; Dordrecht
• Subjects: Biological and medical sciences; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Immunobiology; Myeloid cells: ontogeny, maturation, markers, receptors; Polynuclears; Human; Membrane fraction; Isolation; Chemical composition; Biomembrane; Method; Neutrophil
• ISSN: 1381-5741; 1573-0603
• DOI: 10.1023/A:1009711726462

The surface composition of the azurophilic granule of human neutrophils is largely unknown. To understand the role played by the membrane components of these organelles a simple methodology for their disruption and the isolation of suitable amounts of granule membranes is required. The method of disruption of azurophilic granules available up to now consists of seven steps of freezing-thawing and sonication, a method time-consuming when large amount of granule needs to be processed. In this paper we describe a simple, one-step method based on the granule damaging capacity of Percoll which is suitable for the disruption of large amounts of azurophilic granules. The components of granule lysates obtained with this method were resolved on sucrose gradients. Two major bands were obtained, which are shown to be different from each other judging from their content of selective markers.