Preparation of rat liver plasma membranes in a high yield
โ Scribed by Ernest G. Loten; Jane C. Redshaw-Loten
- Publisher
- Elsevier Science
- Year
- 1986
- Tongue
- English
- Weight
- 231 KB
- Volume
- 154
- Category
- Article
- ISSN
- 0003-2697
No coin nor oath required. For personal study only.
โฆ Synopsis
Existing procedures for the preparation of rat liver plasma membranes are time consuming and generally produce low yields. A method is described in which a rat liver homogenate low-speed pellet is fractionated on a self-forming Percoll gradient. Plasma membranes can be removed from the gradient in a high yield along with much of the DNA in the liver homogenate. A second Percoll step performed in the presence of a low concentration of calcium ions separates the DNA from the plasma membranes. The final membrane fraction has high specific activities of marker enzymes with little contamination with microsomal, mitochondrial, Golgi, or lysosomal markers.
๐ SIMILAR VOLUMES
Further modification of Neville's method of preparation of rat liver plasma membrane has been made in order to study glucagon-sensitive adenylate cyclase. This moditied method introduces two additional steps to the Neville procedures. One involves saving an intermediate layer above the pellet follow
## Identification of gamma-glutamyltransferase in rat liver plasma membranes after two-dimensional electrophoresis Two-dimensional electrophoresis and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, followed by silver staining or specific immunoblotting analysis, was applied to the char