Alexa flour 647 goat anti rabbit igg secondary antibody

Cryopreserved biopsies of patients with Crohn’s disease or with ulcerative colitis embedded in Tissue-Tek O.C.T. compound (Sakura) were acquired from the Basel inflammatory bowel disease cohort. Immunohistochemistry was performed on 6-µm sections using a polyclonal rabbit anti-human CD98hc (ThermoFisher). Primary antibody (1:50 dilution) binding was detected with an Alexa Flour 647 goat anti-rabbit IgG secondary antibody (1:200) (ThermoFisher). Six-micrometer sections from cryopreserved mouse tissues were fixed in 4% paraformaldehyde for 15 min at room temperature. Afterward, sections were blocked with goat serum in DPBS/0.4% Triton-X-100 (Roth) for 30 min and stained with the primary monoclonal rabbit anti-mouse CD98hc mAb overnight in a humidified container at 4 °C. Primary antibody (1:50) binding was detected with an Alexa Flour 647 goat anti-rabbit IgG secondary antibody (1:200) (ThermoFisher). Sections were counterstained with NucBlue^TM Live Cell Stain Ready Probes^TM reagent (1 drop of stain in 500 µl 1× PBS) (Molecular Probes, Hoechst 33342 unique formulation) and imaged with a Nikon A1R Nala confocal microscope.