| Chapter title |
Isolation of Mitochondria from Cultured Cells and Liver Tissue Biopsies for Molecular and Biochemical Analyses
|
|---|---|
| Chapter number | 8 |
| Book title |
Proteomic Profiling
|
| Published in |
Methods in molecular biology, January 2015
|
| DOI | 10.1007/978-1-4939-2550-6_8 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-2549-0, 978-1-4939-2550-6
|
| Authors |
Sabine Schmitt, Carola Eberhagen, Susanne Weber, Michaela Aichler, Hans Zischka |
| Abstract |
We recently reported a new method to isolate functionally intact mitochondria from cell culture and small tissue samples (Schmitt et al., Anal Biochem 443(1):66-74, 2013). This method comprises a semi-automated cell rupture, termed pump controlled cell rupture system (PCC), which can be precisely adjusted to the specific cellular source of isolation and which can be tightly controlled (Schmitt et al., Anal Biochem 443(1):66-74, 2013). Here we provide a detailed hands-on protocol of this PCC method which results in an efficient cell breakage but preserving the mitochondrial integrity. Upon subsequent purification steps, the obtained mitochondrial fraction meets the quality and purity required for molecular analyses, e.g. proteomic comparisons, as well as for biochemical analyses, e.g. determination of diverse enzymatic activities. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Germany | 1 | 5% |
| Unknown | 20 | 95% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Master | 4 | 19% |
| Student > Ph. D. Student | 4 | 19% |
| Professor > Associate Professor | 2 | 10% |
| Student > Bachelor | 2 | 10% |
| Researcher | 2 | 10% |
| Other | 2 | 10% |
| Unknown | 5 | 24% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 8 | 38% |
| Biochemistry, Genetics and Molecular Biology | 2 | 10% |
| Neuroscience | 2 | 10% |
| Computer Science | 1 | 5% |
| Pharmacology, Toxicology and Pharmaceutical Science | 1 | 5% |
| Other | 2 | 10% |
| Unknown | 5 | 24% |