Fluorescein isothiocyanate fitc conjugated anti mouse igg

For immunofluorescence staining [6 (link),11 (link)], cells on glass coverslips or chamber slides were fixed with 4% paraformaldehyde and permeabilized with 0.5% Triton X-100. After washing three times for 10 min with PBS, fixed cells or slides were blocked with blocking buffer (PBS, 0.1% Tween-20, with 3% goat serum) for 1 h and incubated with primary antibody diluted in blocking buffer overnight at 4°C. After washing three times with PBS for 10 min, the fixed cells were treated with the appropriate secondary antibody (fluorescein isothiocyanate (FITC)-conjugated anti-mouse IgG or rhodamine-conjugated anti-rabbit IgG) (Sigma) that was diluted in blocking buffer for 1 h at room temperature. Finally, the fixed cells were washed three times for 10 min with TBS, and their nuclei were counterstained, mounted, and observed by using fluorescence microscope or confocal microscope.