Alexa fluor 555 secondary antibody

Decellularized and control organ samples were fixed in 10% formalin for 24 hours and embedded in paraffin wax for sectioning. Tissue samples were sliced into 4 µm thick sections, heat-fixed and xylene-treated prior to staining. For cellular staining, sections were stained for 10 minutes with Gill III Hematoxylin and Eosin Y (Leica). For immunofluorescence staining, sections were treated with boiling 0.01 M sodium citrate solution supplemented with 0.05% Tween-20 and brought to a pH of 6.0 for antigen retrieval. Sections were blocked with a 10% normal goat serum solution and stained with a 1:200 dilution of myeloperoxidase antibody (Abcam) and 1:500 dilution of Alexa Fluor 488 secondary antibody (Abcam). Tissues were co-stained with a 1:100 dilution of collagen IV or fibronectin antibody (Santa Cruz Biotechnology) and a 1:200 dilution of Alexa Fluor 555 secondary antibody (Santa Cruz Biotechnology). Stained sections were preserved with Vectashield DAPI mounting medium (Vector Labs) and imaged using a Zeiss upright fluorescence microscope. Fluorescence exposure was adjusted as needed to avoid over-saturated images of tissue samples.