An optimized protocol to detect protein ubiquitination and activation by ubiquitination assay in vivo and CCK-8 assay.
STAR Protoc
; 4(2): 102199, 2023 Mar 24.
Article
in En
| MEDLINE
| ID: mdl-36964909
ABSTRACT
E3 ubiquitin ligases play a role in protein degradation, cellular localization, and activation, and their dysregulation is associated with human diseases. Here, we present a protocol to detect IGF2BP1 ubiquitination and activation by an E3 ubiquitin ligase FBXO45. We describe steps for preparing cells and transfecting plasmids. We detail the use of western blot to detect IGF2BP1 ubiquitination and a Cell Counting Kit-8 (CCK-8) assay to detect IGF2BP1 activation. This protocol is applicable to other proteins of interest. For complete details on the use and execution of this protocol, please refer to Lin et al. (2021).1.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Language:
En
Journal:
STAR Protoc
Year:
2023
Document type:
Article
Affiliation country:
China