3 µg of Anti-CD8a-APC or anti-CD8a-PE (clone: 53-6.7 from eBioscience) or purified rabbit anti-mouse collagen IV (Novus Biologicals) were injected intravenously (i.v.). Three minutes later, the animals were sacrificed, lavaged to remove cells in the airway, bled, and perfused with 10 ml of cold PBS. The spleen, LNs, lung, liver, and small intestine were harvested within 12min, and lymphocytes were isolated as described (19 (link)). Immunofluorescence staining was performed as described (6 (link)).
Protocol detail
In Vivo Immunofluorescence Staining
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Corresponding Organization : University of Minnesota
Other organizations : UConn Health
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Variable analysis
independent variables
- Anti-CD8a-APC or anti-CD8a-PE (clone: 53-6.7 from eBioscience) or purified rabbit anti-mouse collagen IV (Novus Biologicals)
- Lymphocytes isolated from the spleen, LNs, lung, liver, and small intestine
- Time between injection and sacrifice (3 minutes)
- Volume of PBS used for perfusion (10 ml)
- Time for tissue harvesting (within 12 minutes)
- No positive or negative controls were explicitly mentioned in the provided information.
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