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A Recombinant Antibody For Tracking Murine Gammaherpesvirus 68 Uracil DNA Glycosylase Expression
Antibodies are powerful tools to detect expressed proteins. However off-target recognition can confound their use. Therefore, careful characterization is needed to validate specificity in distinct applications. Here we report the sequence and characterization of a mouse recombinant antibody that spe...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10245680/ https://www.ncbi.nlm.nih.gov/pubmed/37293087 http://dx.doi.org/10.1101/2023.05.17.541089 |
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author | Mu, Yunxiang Plummer, Joshua B. Zelazowska, Monika A. Paul, Somnath Dong, Qiwen Chen, Zaowen Krug, Laurie T. McBride, Kevin M. |
author_facet | Mu, Yunxiang Plummer, Joshua B. Zelazowska, Monika A. Paul, Somnath Dong, Qiwen Chen, Zaowen Krug, Laurie T. McBride, Kevin M. |
author_sort | Mu, Yunxiang |
collection | PubMed |
description | Antibodies are powerful tools to detect expressed proteins. However off-target recognition can confound their use. Therefore, careful characterization is needed to validate specificity in distinct applications. Here we report the sequence and characterization of a mouse recombinant antibody that specifically detects ORF46 of murine gammaherpesvirus 68 (MHV68). This ORF encodes the viral uracil DNA glycosylase (vUNG). The antibody does not recognize murine uracil DNA glycosylase and is useful in detecting vUNG expressed in virally infected cells. It can detect expressed vUNG in cells via immunostaining and microscopy or flow cytometry analysis. The antibody can detect vUNG from lysates of expressing cells via immunoblot under native conditions but not denaturing conditions. This suggests it recognizes a confirmational based epitope. Altogether this manuscript describes the utility of the anti-vUNG antibody and suitability for use in studies of MHV68 infected cells. |
format | Online Article Text |
id | pubmed-10245680 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Cold Spring Harbor Laboratory |
record_format | MEDLINE/PubMed |
spelling | pubmed-102456802023-06-08 A Recombinant Antibody For Tracking Murine Gammaherpesvirus 68 Uracil DNA Glycosylase Expression Mu, Yunxiang Plummer, Joshua B. Zelazowska, Monika A. Paul, Somnath Dong, Qiwen Chen, Zaowen Krug, Laurie T. McBride, Kevin M. bioRxiv Article Antibodies are powerful tools to detect expressed proteins. However off-target recognition can confound their use. Therefore, careful characterization is needed to validate specificity in distinct applications. Here we report the sequence and characterization of a mouse recombinant antibody that specifically detects ORF46 of murine gammaherpesvirus 68 (MHV68). This ORF encodes the viral uracil DNA glycosylase (vUNG). The antibody does not recognize murine uracil DNA glycosylase and is useful in detecting vUNG expressed in virally infected cells. It can detect expressed vUNG in cells via immunostaining and microscopy or flow cytometry analysis. The antibody can detect vUNG from lysates of expressing cells via immunoblot under native conditions but not denaturing conditions. This suggests it recognizes a confirmational based epitope. Altogether this manuscript describes the utility of the anti-vUNG antibody and suitability for use in studies of MHV68 infected cells. Cold Spring Harbor Laboratory 2023-05-18 /pmc/articles/PMC10245680/ /pubmed/37293087 http://dx.doi.org/10.1101/2023.05.17.541089 Text en https://creativecommons.org/licenses/by-nc/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License (https://creativecommons.org/licenses/by-nc/4.0/) , which allows reusers to distribute, remix, adapt, and build upon the material in any medium or format for noncommercial purposes only, and only so long as attribution is given to the creator. |
spellingShingle | Article Mu, Yunxiang Plummer, Joshua B. Zelazowska, Monika A. Paul, Somnath Dong, Qiwen Chen, Zaowen Krug, Laurie T. McBride, Kevin M. A Recombinant Antibody For Tracking Murine Gammaherpesvirus 68 Uracil DNA Glycosylase Expression |
title | A Recombinant Antibody For Tracking Murine Gammaherpesvirus 68 Uracil DNA Glycosylase Expression |
title_full | A Recombinant Antibody For Tracking Murine Gammaherpesvirus 68 Uracil DNA Glycosylase Expression |
title_fullStr | A Recombinant Antibody For Tracking Murine Gammaherpesvirus 68 Uracil DNA Glycosylase Expression |
title_full_unstemmed | A Recombinant Antibody For Tracking Murine Gammaherpesvirus 68 Uracil DNA Glycosylase Expression |
title_short | A Recombinant Antibody For Tracking Murine Gammaherpesvirus 68 Uracil DNA Glycosylase Expression |
title_sort | recombinant antibody for tracking murine gammaherpesvirus 68 uracil dna glycosylase expression |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10245680/ https://www.ncbi.nlm.nih.gov/pubmed/37293087 http://dx.doi.org/10.1101/2023.05.17.541089 |
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