Autor: |
Susanne Zellner, Karsten Nalbach, Christian Behrends |
Jazyk: |
angličtina |
Rok vydání: |
2021 |
Předmět: |
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Zdroj: |
STAR Protocols, Vol 2, Iss 2, Pp 100506- (2021) |
Druh dokumentu: |
article |
ISSN: |
2666-1667 |
DOI: |
10.1016/j.xpro.2021.100506 |
Popis: |
Summary: 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.For complete details on the use and execution of this protocol, please refer to Zellner et al. (2021). |
Databáze: |
Directory of Open Access Journals |
Externí odkaz: |
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