Anti rabbit igg tritc

hADSCs were differentiated in 24-well chambers at 29 days, rinsed three times with PBS, and incubated in 4% paraformaldehyde overnight at 4°C. Bovine serum albumin (1%) in PBS was used for blocking. Afterwards, the cells were incubated overnight at 4°C with the following primary antibodies: mouse anti-human Cav-1 (1:100; Cell Signaling Technology), rabbit anti-human tyrosine hydroxylase (TH) (1:100; Abcam), rabbit anti-human Lmx1a (1:100; Abcam) or rabbit anti-human Nurr1 (1:100; Santa Cruz Biotechnology). The samples were then rinsed three times thoroughly with PBS and incubated with the following secondary antibodies: anti-rabbit IgG-FITC (Sigma-Aldrich), anti-mouse IgG-FITC (Abcam), anti-mouse IgG-TRITC (Sigma-Aldrich) or anti-rabbit IgG-TRITC (Abcam) for 1.5 hours at room temperature. The cells were thereafter rinsed three times in PBS and incubated for 5 minutes with Hoechst 33258. The samples were then washed twice with PBS and once in deionized water. Stained cells were observed under a confocal laser scanning microscope (SP8, Leica).