Cy2 conjugated goat anti mouse igg

COS7 cells were transfected with 6µg pcDNA-TIRC7-myc and pcDNA3-myc as control. After transfection the expression of recombinant TIRC7 protein was analyzed in transfected COS7 with pcDNA3-TIRC7-myc (COS7^t) and untransfected (COS 7nt) with anti-TIRC7 mAb17. Immunostaining with anti-myc antibody (myc-Ab) (Oncogene) was used as positive control for the transfection. Cells were incubated overnight at 4°C with anti-TIRC7 mAb (10µg/ml) and anti-myc mAb´s (Oncogene,1/10) rinsed with PBS, and incubated with Cy2-conjugated goat anti-mouse IgG (Dianova,1/250) or goat anti-human IgG (Sigma, 1/100) diluted in 5% milk/PBS. Cells were mounted with 50% (v/v) glycerol/PBS and analyzed by confocal laser scanning microscopy.
Wild types and TIRC7 knockout mice splenocytes were fixed for 20 min at 4°C with 4% paraformaldehyde in PBS and permeabilized. Cells were incubated overnight at 4°C with anti-TIRC7 antibody (10µg/ml) or with anti-human IgG (hIgG, 5µg/ml, Sigma) as negative control. Cells were rinsed with PBS, and incubated with Cy3-conjugated goat anti-human IgG (1:250, Sigma) diluted in 5% milk/PBS. Cells were mounted with 50% (vol/vol) glycerol in PBS and confocal images were obtained by using Zeiss LSM 510 confocal laser scanning microscope.

Cells were infected with SARS-CoV-2 and fixed after 20 or 30 h of infection using 4% (w/v) paraformaldehyde/PBS for > 2 h before they were discharged from the BSL-3 laboratory. Cells were permeabilized with 1% (v/v) Triton-X-100/PBS and blocked with 3% (v/v) FCS/PBS. SARS-CoV-2 infection was visualized by use of α-S mAb (kindly provided by Peter Miethe, fzmb, Bad Langensalza, Germany) and Cy2-conjugated goat anti-mouse IgG (Dianova). Nuclei were counterstained with 4′6-diamidino-2-phenylindole (DAPI; Sigma). Fluorescence was visualized using a THUNDER Imager 3D Cell Culture (Leica). Image analysis and processing were performed with LAS X Premium imaging software (Leica).