Optimization of blocking conditions for fluorescent Western blot.
Autofluorescence
Blocking
NF1
Tween 20
Western blot
Journal
Analytical biochemistry
ISSN: 1096-0309
Titre abrégé: Anal Biochem
Pays: United States
ID NLM: 0370535
Informations de publication
Date de publication:
15 03 2020
15 03 2020
Historique:
received:
26
07
2019
revised:
13
01
2020
accepted:
23
01
2020
pubmed:
6
2
2020
medline:
13
1
2021
entrez:
5
2
2020
Statut:
ppublish
Résumé
Blocking conditions are critical for reducing background and non-specific signals in Western blot (WB). For fluorescent WB, however, blockers may bring in additional autofluorescence, and optimal blocking conditions have been less well established. Here, the autofluorescence of Tween 20 is clarified, which is negligible when imaged wet, but could be significant when Tween 20 is dried on blots. In most situations, blocking with Tween 20 reduces background and/or improves specific signals. Systematical investigation of blocking conditions for fluorescent WB reveals that the autofluorescent background is determined by a combination of multiple factors including the blocker, the excitation/emission configuration, the membrane, and the H
Identifiants
pubmed: 32014414
pii: S0003-2697(19)30738-9
doi: 10.1016/j.ab.2020.113598
pii:
doi:
Substances chimiques
Polysorbates
0
Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
113598Informations de copyright
Copyright © 2020 Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of competing interest None.