Anti cd169 clone moma 1

Organ samples were embedded in TissueTek O.T.C. (Sakura) medium and snap frozen in liquid nitrogen. 8 µm sections were generated. Sections were fixed with acetone and unspecific binding sites were blocked After 48 h using two percent fetal calf serum (FCS, Gibco) in PBS. VSV-Glycoprotein (VSV-G) was stained with Anti-VSV-G tag antibody (Abcam). Ebola virus glycoprotein was stained with Anti-Ebola surface glycoprotein (clone KZ52) from absolute antibodies. Secondary antibodies were obtained from ThermoFisher (anti-rabbit). Anti-CD169 (clone MOMA-1) was obtained from Bio-Rad. Antibodies were used in a dilution of 1:100 in blocking buffer and incubated for 30-60 minutes in a humidified darkened chamber at room temperature. Slides were covered with Fluorescence Mounting Medium (Dako, Glostrup, Denkmark) and processed for imaging with a Keyence BZ-9000 microscope.

Histological analysis was performed by preparing 8 µm thick sections from samples of snap frozen organs. IAV-Hemagglutinins were stained with anti-IAV-PR8 Hemagglutinin antibody (Sinobiological). Anti-CD169 (clone MOMA-1) was obtained from Bio-Rad. Anti-CD11c (clone N418) was obtained from eBioscience. Biotinylated Maackia Amurensis Lectin II (MAL II) was purchased from VectorLabs (B-1265-1) and visualized by a secondary Steptavidin-APC antibody staining (Biolegend, 405207).
Acetone (HoneyWell) fixation occurred for 10 min and non-specific binding site blocking was performed using two percent fetal calf serum (FCS, Gibco) in PBS for 15 min. Sections were incubated with antibodies diluted 1:100 in blocking buffer. Antibodies were incubated with the slides for 30–60 min in a humidified darkened chamber. Slides were covered with Fluorescence Mounting Medium (Dako) and image processing was performed using the Keyence BZ-9000 microscope.