Biotin streptavidin conjugated goat anti armenian hamster igg h l

Liver paraffin sections from 12 male mice (3 animals per age group and genotype) were labelled. Antigen retrieval was done by microwaving the sections 4x 5 min in 0.01 M citrate buffer (pH = 6) with 0.05% Tween. All antibodies were diluted in 2% BSA in PBS. Endogenous peroxidase was quenched in 3% H₂O₂ in methanol, and endogenous biotin with Biotin-Blocking System (Agilent, Cat. No X0590). Sections were incubated overnight at 4°C with Armenian hamster anti-mouse CD54 (ICAM-1) (5 μg/ml; Invitrogen—Thermo Fisher Scientific, Cat. No MA5405) or Armenian hamster IgG isotype control (5 μg/ml; Invitrogen—Thermo Fisher Scientific, Cat. No 14-4888-81). After rinsing in PBS with 0.05% Tween, sections were incubated for 30 min at RT with biotin-streptavidin-conjugated goat anti-Armenian hamster IgG (H+L, eBioscience, Cat. No 13–411385; 1.25 μg/ml), washed and incubated for 30 min with Streptavidin peroxidase (Agilent, Cat. No P0397). A diaminobenzidine substrate chromogen system kit (BD Pharmingen, Cat. No 550880) was used to visualize a positive reaction. Sections were counter-stained with haematoxylin, and images taken in a Nikon Eclipse TE2000-U Inverted Microscope.

Liver paraffin sections from 12 male mice (3 animals per age group and genotype) were labelled. Antigen retrieval was done by microwaving the sections 4x 5 min in 0.01 M citrate buffer (pH = 6) with 0.05% Tween. All antibodies were diluted in 2% BSA in PBS. Endogenous peroxidase was quenched in 3% H₂O₂ in methanol, and endogenous biotin with Biotin-Blocking System (Agilent, Cat. No X0590). Sections were incubated overnight at 4°C with Armenian hamster anti-mouse CD54 (ICAM-1) (5 μg/ml; Invitrogen—Thermo Fisher Scientific, Cat. No MA5405) or Armenian hamster IgG isotype control (5 μg/ml; Invitrogen—Thermo Fisher Scientific, Cat. No 14-4888-81). After rinsing in PBS with 0.05% Tween, sections were incubated for 30 min at RT with biotin-streptavidin-conjugated goat anti-Armenian hamster IgG (H+L, eBioscience, Cat. No 13–411385; 1.25 μg/ml), washed and incubated for 30 min with Streptavidin peroxidase (Agilent, Cat. No P0397). A diaminobenzidine substrate chromogen system kit (BD Pharmingen, Cat. No 550880) was used to visualize a positive reaction. Sections were counter-stained with haematoxylin, and images taken in a Nikon Eclipse TE2000-U Inverted Microscope.