The presence of electrical charges on the surface of an organelle is the source of the organelle's electrophoretic mobility. Recently, we reported that capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) can be used to determine the electrophoretic mobility of individual mitochondria. Here, we describe the use of CE-LIF to monitor changes in the electrophoretic mobility distributions of: (i) mitochondria isolated from cultured NS-1 mouse hybridoma cells disrupted by nitrogen cavitation or mechanical homogenization; (ii) mitochondria isolated from rat liver and purified by gradient centrifugation before and after being frozen in liquid nitrogen; and (iii) mitochondria chemically transformed into mitoplasts. These results indicate that the organelle electrophoretic mobility observed by researchers is affected by preparation procedures and that CE-LIF is a complementary technique for monitoring the quality of mitochondrial preparations.
|Number of pages
|Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
|Published - Jul 5 2004
Bibliographical noteFunding Information:
This work was supported by NIH R01-GM61969 (EA). KF was supported by an NIH Chemistry/Biology Interface Training Grant (GM08700). We thank Virginia Haynes and Cecilia Giulivi (Department of Chemistry, University of Minnesota, Duluth) for providing rat liver mitochondria, Sally Palm (Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis) for providing the NS-1 cell line, and Bobby Superfly Poe for obtaining the supplementary electron microscopy images.
- Electrophoretic mobility
- Mitochondrial preparations