Goat anti rabbit igg alexa flour 488

HEK293T or VeroE6-APN cells were grown on coverslips in 6-well plates. At 24 hpt (or h post infection; hpi), cells were washed with phosphate-buffered saline (PBS) and fixed with 4% paraformaldehyde for 20 min at 4 °C. After incubation, cells were washed three times with PBS and blocked with PBS containing 10% FBS, 1% bovine serum albumin (BSA) and 0.2% TritonX-100 for 1 h. Cells were subsequently incubated for 1 h with rabbit anti-myc antibodies (Abcam) and mouse anti-calreticulin ER marker (Abcam) or mouse anti-58K Golgi protein antibodies (Abcam) in 10% FBS at a dilution 1:500 and 1:250, respectively. After washing thrice with PBST, goat anti-rabbit IgG Alexa flour 488 (Abcam) and goat anti-mouse IgG Alexa flour 647 antibodies (Abcam) in 10% FBS at a dilution 1:1000 was added and further incubated for 1 h. The glass slips were mounted on slides with Prolong Gold Antifade Mountant with DAPI (Invitrogen, Carlsbad, CA, USA). The samples were analyzed by FluoviewTM FV1000 confocal microscopy (Olympus, Tokyo, Japan). The Pearson’s correlation coefficients (PCC, Pasadena, CA, USA) were employed to determine co-localization using the ImageJ analysis program [9 (link)] and the PSC co-localization plug-in as described elsewhere [10 (link),11 (link),12 (link)]. Mean Pearson’s coefficient values were calculated from at least ten independent acquired images.