Protocol for immunodot blot detection of UVB photoproducts in extracellular DNA
Summary: UV radiation induces the formation of adducts in genomic DNA within cells that are later found to be present in cell-free fractions associated with extracellular vesicles (EVs) outside of cells. Here, we present a protocol for isolating UV photoproducts in extracellular DNA released from UV...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2024-03-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166724000030 |
| _version_ | 1797350526067671040 |
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| author | M. Alexandra Carpenter Sri Yerrapragada Aleena Alex Michael G. Kemp |
| author_facet | M. Alexandra Carpenter Sri Yerrapragada Aleena Alex Michael G. Kemp |
| author_sort | M. Alexandra Carpenter |
| collection | DOAJ |
| description | Summary: UV radiation induces the formation of adducts in genomic DNA within cells that are later found to be present in cell-free fractions associated with extracellular vesicles (EVs) outside of cells. Here, we present a protocol for isolating UV photoproducts in extracellular DNA released from UVB-irradiated cells via differential centrifugation. We then detail steps for monitoring the DNA adducts using DNA immunoblotting. This protocol can be applied for detection of DNA adducts in EVs from cell culture and skin explant models.For complete details on the use and execution of this protocol, please refer to Carpenter et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-08T12:47:11Z |
| format | Article |
| id | doaj.art-3d742588874d4152aaf02100ace7ef32 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-08T12:47:11Z |
| publishDate | 2024-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-3d742588874d4152aaf02100ace7ef322024-01-21T05:10:38ZengElsevierSTAR Protocols2666-16672024-03-0151102838Protocol for immunodot blot detection of UVB photoproducts in extracellular DNAM. Alexandra Carpenter0Sri Yerrapragada1Aleena Alex2Michael G. Kemp3Department of Pharmacology & Toxicology, Wright State University Boonshoft School of Medicine, Dayton, OH 45435, USA; Corresponding authorDepartment of Pharmacology & Toxicology, Wright State University Boonshoft School of Medicine, Dayton, OH 45435, USADepartment of Pharmacology & Toxicology, Wright State University Boonshoft School of Medicine, Dayton, OH 45435, USADepartment of Pharmacology & Toxicology, Wright State University Boonshoft School of Medicine, Dayton, OH 45435, USA; Dayton VA Medical Center, Dayton, OH 45428, USA; Corresponding authorSummary: UV radiation induces the formation of adducts in genomic DNA within cells that are later found to be present in cell-free fractions associated with extracellular vesicles (EVs) outside of cells. Here, we present a protocol for isolating UV photoproducts in extracellular DNA released from UVB-irradiated cells via differential centrifugation. We then detail steps for monitoring the DNA adducts using DNA immunoblotting. This protocol can be applied for detection of DNA adducts in EVs from cell culture and skin explant models.For complete details on the use and execution of this protocol, please refer to Carpenter et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166724000030Cell cultureCell isolationCell-based AssaysCell separation/fractionationAntibodySignal Transduction |
| spellingShingle | M. Alexandra Carpenter Sri Yerrapragada Aleena Alex Michael G. Kemp Protocol for immunodot blot detection of UVB photoproducts in extracellular DNA STAR Protocols Cell culture Cell isolation Cell-based Assays Cell separation/fractionation Antibody Signal Transduction |
| title | Protocol for immunodot blot detection of UVB photoproducts in extracellular DNA |
| title_full | Protocol for immunodot blot detection of UVB photoproducts in extracellular DNA |
| title_fullStr | Protocol for immunodot blot detection of UVB photoproducts in extracellular DNA |
| title_full_unstemmed | Protocol for immunodot blot detection of UVB photoproducts in extracellular DNA |
| title_short | Protocol for immunodot blot detection of UVB photoproducts in extracellular DNA |
| title_sort | protocol for immunodot blot detection of uvb photoproducts in extracellular dna |
| topic | Cell culture Cell isolation Cell-based Assays Cell separation/fractionation Antibody Signal Transduction |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724000030 |
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