Inactivation of Horseradish Peroxidase by Acid for Sequential Chemiluminescent Western Blot.
acetic acid
horseradish peroxidase
hydrogen peroxide
western blot
Journal
Biotechnology journal
ISSN: 1860-7314
Titre abrégé: Biotechnol J
Pays: Germany
ID NLM: 101265833
Informations de publication
Date de publication:
Mar 2020
Mar 2020
Historique:
received:
04
09
2019
pubmed:
28
10
2019
medline:
29
9
2020
entrez:
25
10
2019
Statut:
ppublish
Résumé
Chemiluminescent western blot (WB) is often performed sequentially for detection of overlapping proteins; in between, prior antibodies must be stripped or the conjugated horseradish peroxidase (HRP) inactivated. However, often, stripping either is insufficient to remove all the bound antibodies or causes protein loss, whereas treatment with hydrogen peroxide, a popular way to inactivate HRP, may affect epitope recognition as the authors previously reported. To date, an ideal method for sequential chemiluminescent WB is still missing. Here it is demonstrated that acid equivalent to 10% acetic acid can efficiently inactivate HRP, allowing sequential probing without protein loss or epitope damage.
Identifiants
pubmed: 31647189
doi: 10.1002/biot.201900397
doi:
Substances chimiques
Hydrogen Peroxide
BBX060AN9V
Horseradish Peroxidase
EC 1.11.1.-
Acetic Acid
Q40Q9N063P
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e1900397Subventions
Organisme : Deutsche Forschungsgemeinschaft
ID : MO 1421/5-1
Informations de copyright
© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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