Immunocytochemistry analysis of SiHa cells labeling CYP7A1 with Rabbit anti-CYP7A1 antibody (HA500207) at 1/200 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-CYP7A1 antibody (HA500207) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) were used as the secondary antibody at 1/1,000 dilution.

Flow cytometric analysis of CYP7A1 was done on HeLa cells. The cells were fixed, permeabilized and stained with the primary antibody (HA500207, 1ug/ml) (red) compared with Rabbit IgG, monoclonal - Isotype Control (green). After incubation of the primary antibody at +4℃ for 1 hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃ (dark incubation).Unlabelled sample was used as a control (cells without incubation with primary antibody; black).