Work

Organelle Isolation by Magnetic Immunoabsorption

Year: 1999

Type: article

Abstract: Currently, most organelle isolation procedures rely on physical parameters and centrifugation for separation. Here, we report the rapid and gentle isolation of a variety of organelles by immunolabeling whole cell lysates with organelle-specific antibodies and streptavidin magnetic particles followed by separation in a magnetic field. Using magnetic immunoabsorption, we have been able to specifically label mouse metaphase chromosomes and a variety of plant organelles, including: amyloplasts, choroplasts and nuclei from whole cell lysates of various plant tissues. We find that the distinct magnetic properties, surface characteristics and mean diameter-size ranges of different particle preparations significantly influence their specific utility for organelle isolations. By using an internal-field magnetic separation device, we have developed a method for quantitative recovery of labeled organelles in microarrays and tested a variety of antibodies to chloroplast outer envelope proteins for their ability to immune-isolate chloroplasts.

Source: BioTechniques

DOI: 10.2144/99262rr04

Authors: Albert P. Kausch, T. Page Owen, S. Narayanswami, Barry D. Bruce

Institutions: University of Connecticut, Connecticut College, Jackson Laboratory, University of Tennessee at Knoxville

Language: English

Topic: Biotin and Related Studies

Subfield: Cell Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $1,415

OpenAlex ID: W117701556

FWCI: 0.62

Cites: 9

Cited by: 31

Related to: 10