Lymph nodes were fixed with 4% PFA for two hours, incubated in 30% sucrose, and frozen in O.C.T. Lungs were inflated with, then incubated in 4% PFA for 3 hours, and incubated in 30% sucrose prior to freezing. Eight micron sections were cut and epitopes on slides were stained using tyramide amplification as described 21 (link). eGFP and YFP were detected using a purified polyclonal rabbit anti-mouse GFP (NB600-308; Novus Biologicals) followed by biotinylated F(ab')2 donkey anti-rabbit (Jackson ImmunoResearch Laboratories). Biotinylated anti-mouse CD4 (RM4-5; Biolegend); IgD (11-26c; eBiosciences) and FDCM2 (212-MK-2FDCM2; ImmunoKontact) were used to detect CD4 T cells, B cells, and FDC, respectively. 4′,6-Diamidine-2′-phenylindole dihydrochloride (DAPI; 0.5μg/ml; Roche) in PBS was used to counterstain nuclei prior to mounting on coverslips with Vectashield (Vector laboratories). Images were collected with a Zeiss Axio Imager. Analysis of images was performed using FIJI (ImageJ) software.
Biotinylated f ab 2 donkey anti rabbit
Manufactured by Jackson ImmunoResearch
Biotinylated F(ab')2 donkey anti-rabbit is a laboratory reagent produced by Jackson ImmunoResearch. It consists of the F(ab')2 fragment of donkey-derived antibodies that have been labeled with biotin.
Automatically generated - may contain errors
Lab products found in correlation
Axio imager, by Zeiss (2 mentions)
Nb600 308, by Novus Biologicals (2 mentions)
Vectashield, by Vector Laboratories (2 mentions)
Biotinylated anti mouse cd4 rm4 5, by BioLegend (2 mentions)
Igd 11 26c, by Thermo Fisher Scientific (2 mentions)
4 6 diamidine 2 phenylindole dihydrochloride dapi, by Roche (2 mentions)
2 protocols using biotinylated f ab 2 donkey anti rabbit
1
Multicolor Immunostaining of Lymphoid Tissues
2
Multicolor Immunostaining of Lymphoid Tissues
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Lymph nodes were fixed with 4% PFA for two hours, incubated in 30% sucrose, and frozen in O.C.T. Lungs were inflated with, then incubated in 4% PFA for 3 hours, and incubated in 30% sucrose prior to freezing. Eight micron sections were cut and epitopes on slides were stained using tyramide amplification as described 21 (link). eGFP and YFP were detected using a purified polyclonal rabbit anti-mouse GFP (NB600-308; Novus Biologicals) followed by biotinylated F(ab')2 donkey anti-rabbit (Jackson ImmunoResearch Laboratories). Biotinylated anti-mouse CD4 (RM4-5; Biolegend); IgD (11-26c; eBiosciences) and FDCM2 (212-MK-2FDCM2; ImmunoKontact) were used to detect CD4 T cells, B cells, and FDC, respectively. 4′,6-Diamidine-2′-phenylindole dihydrochloride (DAPI; 0.5μg/ml; Roche) in PBS was used to counterstain nuclei prior to mounting on coverslips with Vectashield (Vector laboratories). Images were collected with a Zeiss Axio Imager. Analysis of images was performed using FIJI (ImageJ) software.
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