A Simple, Reproducible Procedure for Chemiluminescent Western Blot Quantification.
Analysis
Data normalization
Linear regression model
Protein
Quantification
Western blot
Journal
Bio-protocol
ISSN: 2331-8325
Titre abrégé: Bio Protoc
Pays: United States
ID NLM: 101635102
Informations de publication
Date de publication:
05 May 2023
05 May 2023
Historique:
received:
10
11
2022
revised:
31
01
2023
accepted:
14
03
2023
medline:
16
6
2023
pubmed:
16
6
2023
entrez:
16
6
2023
Statut:
epublish
Résumé
Western blotting is a universally used technique to identify specific proteins from a heterogeneous and complex mixture. However, there is no clear and common procedure to quantify the results obtained, resulting in variations due to the different software and protocols used in each laboratory. Here, we have developed a procedure based on the increase in chemiluminescent signal to obtain a representative value for each band to be quantified. Images were processed with ImageJ and subsequently compared using R software. The result is a linear regression model in which we use the slope of the signal increase within the combined linear range of detection to compare between samples. This approach allows to quantify and compare protein levels from different conditions in a simple and reproducible way. Graphical overview.
Identifiants
pubmed: 37323629
doi: 10.21769/BioProtoc.4667
pii: e4667
pmc: PMC10266446
doi:
Types de publication
Journal Article
Langues
eng
Pagination
e4667Informations de copyright
Copyright © 2023 The Authors; exclusive licensee Bio-protocol LLC.
Déclaration de conflit d'intérêts
Competing interestsThe authors declare no competing interests and mentioning of specific materials, reagents, and equipment does not imply endorsement by the funding agencies.
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