Chang J, et al. (2008) Purification of yeast membranes and organelles by sucrose density gradient centrifugation. Methods Mol Biol 457:141-9
Abstract: Many experiments require isolation and purification of membranes and organelles from a cell-free lysate. A combination of differential and sucrose density gradient centrifugation provides adequate separation of most yeast organelles in a single experiment. Yeast cells are converted to spheroplasts and gently lysed under conditions that preserve the integrity of organelles. The total lysate is subjected to differential centrifugation and the resulting membrane pellets are fractionated on density gradients. The method is based on the fact that different membranes contain different ratios of lipid to protein, and thus exhibit different density, allowing them to migrate through the gradient until they reach isopycnic position. The fractionated gradients are analyzed by Western blotting with antibodies that recognize marker proteins specific for individual organelles.
|Status: Published||Type: Journal Article||PubMed ID: 19066024|
Topics addressed in this paper
Number of different genes curated to this paper: 3
- To find other papers on a gene and topic, click on the colored ball in the appropriate box.
- displays other papers with information about that topic for that gene.
- displays other papers in SGD that are associated with that topic.
The topic is addressed in these papers but does not describe a specific gene or chromosomal feature.
- To go to the Locus page for a gene, click on the gene name.