Retro-orbital CD45 antibody labeling to evaluate antibody-secreting cell trafficking in mice
Summary: Antibody-secreting cells (ASCs) are critical regulators of the humoral immune response. However, differences between tissue resident populations versus those that have recently migrated to their final anatomic destination are poorly understood. Here, we present a protocol for using retro-or...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2023-06-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723002757 |
| _version_ | 1797827801534955520 |
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| author | KimAnh T. Pioli Peter D. Pioli |
| author_facet | KimAnh T. Pioli Peter D. Pioli |
| author_sort | KimAnh T. Pioli |
| collection | DOAJ |
| description | Summary: Antibody-secreting cells (ASCs) are critical regulators of the humoral immune response. However, differences between tissue resident populations versus those that have recently migrated to their final anatomic destination are poorly understood. Here, we present a protocol for using retro-orbital (r.o.) CD45 antibody labeling to identify tissue resident versus recently immigrated ASCs in mice. We describe steps for r.o. injection of antibodies, animal euthanasia, and tissue harvesting. We then detail tissue processing, cell counting, and cell staining for flow cytometry analysis.For complete details on the use and execution of this protocol, please refer to Pioli et al. (2023).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-09T12:54:11Z |
| format | Article |
| id | doaj.art-c67cfbe83b304b47b4dad6c97c7900a8 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-09T12:54:11Z |
| publishDate | 2023-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-c67cfbe83b304b47b4dad6c97c7900a82023-05-14T04:29:50ZengElsevierSTAR Protocols2666-16672023-06-0142102308Retro-orbital CD45 antibody labeling to evaluate antibody-secreting cell trafficking in miceKimAnh T. Pioli0Peter D. Pioli1Department of Biochemistry, Microbiology and Immunology, College of Medicine, University of Saskatchewan, Saskatoon, SK S7N5E5, Canada; Corresponding authorDepartment of Biochemistry, Microbiology and Immunology, College of Medicine, University of Saskatchewan, Saskatoon, SK S7N5E5, Canada; Corresponding authorSummary: Antibody-secreting cells (ASCs) are critical regulators of the humoral immune response. However, differences between tissue resident populations versus those that have recently migrated to their final anatomic destination are poorly understood. Here, we present a protocol for using retro-orbital (r.o.) CD45 antibody labeling to identify tissue resident versus recently immigrated ASCs in mice. We describe steps for r.o. injection of antibodies, animal euthanasia, and tissue harvesting. We then detail tissue processing, cell counting, and cell staining for flow cytometry analysis.For complete details on the use and execution of this protocol, please refer to Pioli et al. (2023).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/S2666166723002757Flow Cytometry/Mass CytometryImmunologyModel OrganismsAntibody |
| spellingShingle | KimAnh T. Pioli Peter D. Pioli Retro-orbital CD45 antibody labeling to evaluate antibody-secreting cell trafficking in mice STAR Protocols Flow Cytometry/Mass Cytometry Immunology Model Organisms Antibody |
| title | Retro-orbital CD45 antibody labeling to evaluate antibody-secreting cell trafficking in mice |
| title_full | Retro-orbital CD45 antibody labeling to evaluate antibody-secreting cell trafficking in mice |
| title_fullStr | Retro-orbital CD45 antibody labeling to evaluate antibody-secreting cell trafficking in mice |
| title_full_unstemmed | Retro-orbital CD45 antibody labeling to evaluate antibody-secreting cell trafficking in mice |
| title_short | Retro-orbital CD45 antibody labeling to evaluate antibody-secreting cell trafficking in mice |
| title_sort | retro orbital cd45 antibody labeling to evaluate antibody secreting cell trafficking in mice |
| topic | Flow Cytometry/Mass Cytometry Immunology Model Organisms Antibody |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723002757 |
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