論文 - 詳細
| RRC ID | 19219 |
|---|---|
| 著者 | Eng KE, Panas MD, Karlsson Hedestam GB, McInerney GM. |
| タイトル | A novel quantitative flow cytometry-based assay for autophagy. |
| ジャーナル | Autophagy |
| Abstract |
Autophagy is a cellular degradation process with an increasingly recognised importance in many biological pathways such as nutrient sensing, stress responses and development. We present a straightforward assay for autophagy which combines the sensitivity of the EGFP-LC3 reporter protein with the throughput capacity and quantitative power of flow cytometry. Because saponin extraction is specific for the non-autophagosome associated EGFP-LC3-I form of the protein, flow cytometry can be used to measure total fluorescence of saponin extracted HOS-EGFP-LC3 cells as a measure of the levels of autophagosome associated EGFP-LC3-II. Combined with inhibitors of degradation, we have adapted this assay to differentiate between constitutive and induced autophagy and to quantify the changes in flux of the system. Moreover, using direct antibody staining for the endogenous LC3 protein, we have extended this assay to the detection of autophagosome formation in non-transfected cells. |
| 巻・号 | 6(5) |
| ページ | 634-41 |
| 公開日 | 2010-7-1 |
| DOI | 10.4161/auto.6.5.12112 |
| PII | 12112 |
| PMID | 20458170 |
| MeSH | Animals Autophagy* / drug effects Biological Assay / methods* Chloroquine / pharmacology Flow Cytometry / methods* Green Fluorescent Proteins / metabolism Humans Intracellular Space / drug effects Intracellular Space / metabolism Mice Microtubule-Associated Proteins / metabolism Saponins / metabolism |
| IF | 9.77 |
| 引用数 | 98 |
| WOS 分野 | CELL BIOLOGY |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 7 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | Atg5^(+/+)MEF(RCB2710) Atg5^(-/-)MEF(RCB2711) |