| Autor: |
Zellner, Susanne, Nalbach, Karsten, Behrends, Christian |
| Zdroj: |
STAR Protocols; June 2021, Vol. 2 Issue: 2 |
| Abstrakt: |
The ascorbate peroxidase APEX2 is commonly used to study the neighborhood of a protein of interest by proximity-dependent biotinylation. Here, we describe a protocol for sample processing compatible with immunoblotting and mass spectrometry, suitable to specifically map the content of autophagosomes and potentially other short-lived endomembrane transport vesicles without the need of subcellular fractionation. By combining live-cell biotinylation with proteinase K digestion of cell homogenates, proteins enriched in membrane-protected compartments can be readily enriched and identified. |
| Databáze: |
Supplemental Index |
| Externí odkaz: |
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