Rabbit anti horse spleen ferritin antibody

Immunofluorescence was performed on formalin-fixed tissue after MRI and stereological analysis to confirm labeling of CF in the kidney. Tissue samples were embedded in paraffin and sectioned to 4-μm thickness. Sections were rehydrated using Histoclear (National Diagnostics, GA, USA), followed by serially decreasing ethanol dilutions. Sections were then subjected to antigen retrieval, using a 10 mM citrate buffer for 30 min at 100° C. Tissue was blocked with Dako blocking solution for 1 h in a humidity chamber. Sections were labeled first with a rabbit anti-horse spleen ferritin antibody (Sigma Aldrich, St Louis, MO, USA) overnight, then labeled with donkey anti-goat Alexa 488 for 2 h followed by donkey anti-rabbit Alexa 568 (1:500; Life Technologies) for 2 h at room temperature under light protection. 4′,6-diamidino-2-phenylindole (DAPI) was applied to stain nuclei and samples were dehydrated in solutions of serially decreasing ethanol followed by Histoclear. Images were obtained on a Leica confocal microscope (Leica MicroSystems, Manheim, Germany) using a 40x objective.