Site-specific Labeling of B Cell Receptor and Soluble Immunoglobulin.
B lymphocytes
BCR
Conformational changes
Site-specific labeling
Journal
Bio-protocol
ISSN: 2331-8325
Titre abrégé: Bio Protoc
Pays: United States
ID NLM: 101635102
Informations de publication
Date de publication:
20 Sep 2020
20 Sep 2020
Historique:
received:
28
05
2020
revised:
19
07
2020
accepted:
14
08
2020
entrez:
4
3
2021
pubmed:
5
3
2021
medline:
5
3
2021
Statut:
epublish
Résumé
B lymphocyte activation is regulated by its membrane-bound B cell receptors (BCRs) upon recognizing diverse antigens. It is hypothesized that antigen binding would trigger conformational changes within BCRs, followed by a series of downstream signaling activation. To measure the BCR conformational changes in live cells, a fluorescent site-specific labeling technique is preferred. Genetically encoded fluorescent tags visualize the location of the target proteins. However, these fluorescent proteins are large (~30 kDa) and would potentially perturb the conformation of BCRs. Here, we describe the general procedures of utilizing short tag-based site-specific labeling methodologies combining with fluorescence resonance energy transfer (FRET) assay to monitor the conformational changes within BCR extracellular domains upon antigen engagement.
Identifiants
pubmed: 33659425
doi: 10.21769/BioProtoc.3767
pii: e3767
pmc: PMC7842509
doi:
Types de publication
Journal Article
Langues
eng
Pagination
e3767Informations de copyright
Copyright © The Authors; exclusive licensee Bio-protocol LLC.
Déclaration de conflit d'intérêts
Competing interestsThe authors declare no financial or commercial conflict of interest.
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