Ab1422

For flow cytometry the following antibodies were used: PE-conjugated mouse anti- HLA-ABC (1:100, 555553, BD Biosciences), human anti- HLA-A2 antibody (3 µg mL^-1, SN607D8), mouse anti-human CD55 (1:500, ab1422, Abcam), mouse anti-C3 (1:1000, RFK22, in-house generated), mouse anti-C5b-9 (1:100, AE11, Hycult Biotech), PE-conjugated mouse anti-human CD46 (1:100, 12-0469-42, Invitrogen), and APC-conjugated mouse anti-human CD59 (1:100, 17-0596-42, Invitrogen) were used as primary antibodies. Secondary antibodies used for detection were PE-conjugated goat anti-human IgG (1:1000, 109-116-098, Jackson) and PE-conjugated goat anti-mouse (1:100, R0480, DAKO). All antibody incubations were done for 30 min on ice with subsequent washing in PBS + 0,1% BSA. Flow cytometry was performed on a LSR-II (BD) and acquired results analyzed using FlowJo (BD).

Organoids were fixed in 2% PFA for 20 min at 4°C, washed with PBS and incubated with the following primary antibodies: sheep anti-human NPHS1 (1:100, AF4269, R&D Systems), mouse anti-E-Cadherin (1:250, 610181, BD), biotinylated Lotus Tetragonolobus Lectin (LTL) (1:300, B-1325, Vector Laboratories) and mouse anti-human CD55 (1:500, ab1422, Abcam). Secondary antibodies were donkey anti-sheep Alexa-647 (A21448, Invitrogen), donkey anti-mouse Alexa-568 (A10037, Invitrogen), donkey anti-mouse Alexa-488 (A21202, Invitrogen), and Streptavidin conjugated with Alexa-532 (S11224, Invitrogen), all at 1:500 dilution. Nuclei were stained with Hoechst33342 (1:10000, H3576, Invitrogen) and organoids were mounted using ProLong Gold Antifade Mountant (Invitrogen) in glass bottom dishes (MatTek). Images were acquired using a White Light Laser Confocal Microscope TCS SP8 and LAS-X Image software.