labelling endothelial cells  (Novus Biologicals)

Journal: Brain, behavior, & immunity - health

Article Title: Investigation in blood-brain barrier integrity and susceptibility to immune cell penetration in a mouse model of Dravet syndrome.

doi: 10.1016/j.bbih.2025.100955

Figure Lengend Snippet: Fig. 3. Evaluation of neutrophil infiltration into the brain parenchyma: Representative images of a double immunofluorescence of the endothelial marker laminin (red) and neutrophils (NIMP-R14, green) in the hippocampus of Syn1-Cre/Scn1aWT/A1783V mice and their control group (Scn1aWT/WT) at PND25. Positive control corresponds to an ischemic brain tissue with proven infiltration of neutrophils into the brain ischemic core. Scale bar = 50 μm.

Article Snippet: Sections were mounted on gelatin-coated slides and, once adhered, washed with TBS, permeabilized with TBS containing Triton X-100 0.2% for 30 min, and blocked with TBS containing Triton X-100 0.1% and BSA 5% for 1 h. After several washes with TBS, sections were incubated overnight at 4 ◦C with the following primary antibodies: (i) Analysis of neutrophils: antilaminin rabbit polyclonal antibody (ref. NB300-144; Novus Biologicals, Centennial, CO, USA) used at 1/200, and anti-NIMP-R14 rat monoclonal antibody (ref. ab2557; Abcam, Cambridge, UK) used at 1/200 for labelling neutrophils; and (ii) Analysis of B and T cells: antiCD31/ PECAM-1 rat monoclonal antibody (ref. 553370, BD-Biosciences, Madrid, Spain) used at 1/100 for labelling endothelial cells, antiB220/CD45R rabbit monoclonal antibody (ref. NBP3-11980; Novus Biological Centennial, CO, USA) used at 1/250 for labelling B-cells, and anti-CD3 rabbit polyclonal antibody (ref. ab5690, Abcam, Cambridge, UK) used at 1/250 for labelling T-cells.

Techniques: Immunofluorescence, Marker, Control, Positive Control