Tyramide Signal Amplification Kit - 5 Color (A329043)

Four-color TSA immunohistochemistry of PFA-fixed, frozen mouse small intestine section using antibodies that have been biotinylated and labelled with the StreptaClick® HRP Complete Tyramide Signal Amplification and Biotinylation Kit - 5 Color. Tissue was mounted in OCT compound and frozen in liquid nitrogen before cutting 10 µm sections. The frozen tissue section was fixed in acetone (30 seconds in ice cold 50% acetone/50%water + 10 min in ice cold 100% acetone), air dried, and washed in PBS. Immunohistochemistry was performed using the following biotinylated antibodies: biotin rat anti-CD31 [MEC13.3] (Tyramide 488; green), biotin mouse anti-human alpha actin [1A4] (Tyramide 594; red), biotin rat anti-CD326 [G8.8] (Tyramide 555; yellow), biotin rat anti-mouse CD8a [53-6.7] (Tyramide 647; blue).

Five-color TSA immunohistochemistry of PFA-fixed, frozen mouse spleen section using antibodies that have been biotinylated and labelled with the StreptaClick® HRP Complete Tyramide Signal Amplification and Biotinylation Kit - 5 Color. Tissue was fixed in 4% paraformaldehyde/10% sucrose for 30 min at +20°C, followed by incubation in 30% sucrose at +4°C overnight before freezing and cutting 10 µm sections. The frozen tissue section was fixed in acetone at room temperature for 10 min, air dried, and washed in PBS. Immunohistochemistry was performed using biotinylated antibodies and TSA dyes for color development: biotin rat anti-mouse CD4 [GK1.5] (Tyramide 430; cyan), biotin hamster anti-mouse CD11c [N418] (Tyramide 488; green), biotin rat anti-mouse B220 [RA3-6B2] (Tyramide 555; yellow), biotin rat anti-mouse CD8a [53-6.7] (Tyramide 594; red), and biotin rat anti-mouse F4/80 [M1/70] (Tyramide 647; magenta).