Protocol for the quantitative identification of endogenously ISGylated proteins from mammalian cell lines.
STAR Protoc
; 5(1): 102843, 2024 Mar 15.
Article
in En
| MEDLINE
| ID: mdl-38294909
ABSTRACT
Ubiquitin-like protein ISG15 plays an important role in an array of cellular functions via its covalent attachment to target proteins (ISGylation). Here, we present a protocol for the identification of ISGylated proteins that avoids the caveats associated with ISG15 overexpression and minimizes the likelihood of false positives. We describe steps for the tagging of endogenous ISG15, followed by genotyping and clone selection. We then detail steps for ISGylation induction, the isolation of ISGylated proteins, and their identification via quantitative mass spectrometry. For complete details on the use and execution of this protocol, please refer to Wardlaw and Petrini.1.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Ubiquitins
/
Cytokines
Type of study:
Diagnostic_studies
Limits:
Animals
Language:
En
Journal:
STAR Protoc
Year:
2024
Document type:
Article
Country of publication: