Rabbit antibody to TGN38 (310-360). Human Melanoma cell line C 32 was cultured overnight on round cover slides placed in a 24 well tissue culture plate. Culture media removed and washed twice with PBS before fixing with 2% formalin for 10 minutes. Cells were then washed three times with PBS and incubated with Tris 0.01M containing Triton X 0.005% for 15 minutes. Cells were washed and incubated with 100 µl of Rabbit antibody to TGN diluted 1:100 in the blocking buffer for 30 minutes. Wells were then washed 7 times with PBS and incubated with 100 µl of anti Rb-FITC conjugate diluted 1:100 in the blocking buffer for further 30 minutes. Cells were washed as before and nuclear counter stained with Hoechst and mounted on to slides.