Protocol for Efficient Protein Synthesis Detection by Click Chemistry in Colorectal Cancer Patient-Derived Organoids Grown In Vitro.
STAR Protoc
; 1(2): 100103, 2020 09 18.
Article
em En
| MEDLINE
| ID: mdl-33000005
ABSTRACT
Here, we describe a protocol to detect and visualize protein synthesis by click-chemistry-based immunofluorescence in patient-derived organoids (PDOs) in vitro. The protocol uses O-propargyl puromycin (OPP), an analog of puromycin that enters the acceptor site of ribosomes and is incorporated into nascent polypeptides. OPP can be detected by a click chemistry reaction and can be combined with conventional antibody staining. We describe procedures for imaging intact organoids in 3D format or imaging sections of organoids from paraffin blocks. For complete details on the use and execution of this protocol, please refer to Morral, Stanisavljevic et al. (2020).
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Puromicina
/
Proteínas
/
Imunofluorescência
/
Química Click
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article
País de afiliação:
Espanha