Quantification of total and phosphorylated STAT3 by calibrated western blotting.
Antibody
Protein Biochemistry
Signal Transduction
Journal
STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501
Informations de publication
Date de publication:
04 Sep 2023
04 Sep 2023
Historique:
received:
05
05
2023
revised:
23
06
2023
accepted:
25
07
2023
medline:
5
9
2023
pubmed:
5
9
2023
entrez:
5
9
2023
Statut:
aheadofprint
Résumé
Quantification of intracellular proteins is essential to understand signaling. Here, we describe quantification of the expression and phosphorylation of the transcription factor STAT3. We present isolation of total and phosphorylated STAT3 from cell lysates by immunoprecipitation, followed by SDS-PAGE and western blot together with known amounts of a calibrator protein that shares an epitope with the precipitated proteins. Finally, we explain how to relate the amount of precipitated protein to the amount of calibrator protein considering the efficiency of immunoprecipitation. For complete details on the use and execution of this protocol, please refer to Dittrich et al. (2012)
Identifiants
pubmed: 37669163
pii: S2666-1667(23)00475-6
doi: 10.1016/j.xpro.2023.102508
pmc: PMC10485629
pii:
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
102508Informations de copyright
Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of interests The authors declare no competing interests.
Références
Mol Biosyst. 2012 Aug;8(8):2119-34
pubmed: 22711165
Cytokine Growth Factor Rev. 2015 Oct;26(5):595-602
pubmed: 26187858
Cell Commun Signal. 2019 May 17;17(1):46
pubmed: 31101051
STAR Protoc. 2023 Aug 14;4(3):102511
pubmed: 37581983