Single Cell Determination of 7,8-dihydro-8-oxo-2'-deoxyguanosine by Fluorescence Techniques: Antibody vs. Avidin Labeling.
7,8-dihydro-8-oxo-2′-deoxyguanosine
avidin binding
immunofluorescence
oxidative DNA damage
Journal
Molecules (Basel, Switzerland)
ISSN: 1420-3049
Titre abrégé: Molecules
Pays: Switzerland
ID NLM: 100964009
Informations de publication
Date de publication:
25 May 2023
25 May 2023
Historique:
received:
21
03
2023
revised:
11
05
2023
accepted:
23
05
2023
medline:
12
6
2023
pubmed:
10
6
2023
entrez:
10
6
2023
Statut:
epublish
Résumé
An important biomarker of oxidative damage in cellular DNA is the formation of 7,8-dihydro-8-oxo-2'-deoxyguanosine (8-oxodG). Although several methods are available for the biochemical analysis of this molecule, its determination at the single cell level may provide significant advantages when investigating the influence of cell heterogeneity and cell type in the DNA damage response. to. For this purpose, antibodies recognizing 8-oxodG are available; however, detection with the glycoprotein avidin has also been proposed because of a structural similarity between its natural ligand biotin and 8-oxodG. Whether the two procedures are equivalent in terms of reliability and sensitivity is not clear. In this study, we compared the immunofluorescence determination of 8-oxodG in cellular DNA using the monoclonal antibody N45.1 and labeling using avidin conjugated with the fluorochrome Alexa Fluor488 (AF488). Oxidative DNA damage was induced in different cell types by treatment with potassium bromate (KBrO
Identifiants
pubmed: 37298802
pii: molecules28114326
doi: 10.3390/molecules28114326
pmc: PMC10254197
pii:
doi:
Substances chimiques
8-Hydroxy-2'-Deoxyguanosine
88847-89-6
Deoxyguanosine
G9481N71RO
Avidin
1405-69-2
Biomarkers
0
Antibodies, Monoclonal
0
DNA
9007-49-2
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : CNR (Italy)/CONCYTEC (Peru)
ID : Biennial Program 2021-2022
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