Fitc conjugated goat anti mouse igg ab6785

DCs (1×10⁵) were pulsed with 10 µg M-HSP70-PCs purified from the six melanoma cell lines at 37°C for 12 h. After culture, cytospin slides were prepared by centrifugation (100 × g for 5 min) using 100 µl of cell suspension containing about 1×10⁶ cells/ml. The cells were air-dried, immediately fixed with 4% paraformaldehyde, permeabilized with 0.5% Triton X-100, and blocked in 3% bovine serum albumin (BSA) at 4°C for 1 h. The cells were then incubated with monoclonal rabbit anti-Melan-A and monoclonal mouse anti-NY-ESO-1 antibodies at a 1:100 dilution in PBS for 1 h at 37°C. After three washes in PBS, the cells were incubated with Texas Red-conjugated goat anti-rabbit IgG (ab6719) and FITC-conjugated goat anti-mouse IgG (ab6785; both from Abcam) at a 1:50 dilution in PBS for 30 min in the dark at 37°C. The cells were washed again three times in PBS and smeared onto the slides. Images were acquired using a Olympus DP71 microscope (Olympus) and analyzed by the included software.