Dylight 488 conjugated goat anti rabbit igg

Trypsin was purchased from Solarbio (Beijing, China), DNase I was purchased from Roche Diagnostics GmbH (Mannheim, Germany), DMEM/F-12 was purchased from Thermo Fisher Scientific Inc. (Waltham, MA, USA), newborn calf serum was purchased from Shanghai ExCell Biology, Inc. (Shanghai, China), dimethyl sulfoxide was purchased from Biosharp (Hefei, China), chenodeoxycholic acid (CDCA) was purchased from Shanghai Yiji Co., Ltd. (Shanghai, China), Z-guggulsterone, rabbit polyclonal anti-FXR antibody, and DAPI were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and rat IgG and DyLight® 488-conjugated goat anti-rabbit IgG were purchased from EarthOx, LLC (Millbrae, California, USA).

To verify the identity and assess the purity of isolated glial cells, immunofluorescent microscopy was used. Cells plated on poly-D-lysine-coated coverslips were fixed in acetic acid/ethanol [5:95 (v/v)] at −20 °C for 10 min and then incubated with the relevant primary antibody for 30 min at room temperature, followed by three washes in Minimum Essential Medium (MEM) (Invitrogen) and subsequent incubation with secondary antibody for 30 min. Sections were mounted in a medium containing 4′,6′-diamidino-2-phenylindole dihydrochloride (DAPI) (100 ng/mL; Vector Laboratories) to show cell nuclei, and examined by confocal microscopy (Olympus FluoView FV10i; Olympus, Tokyo, Japan). The primary antibodies used were anti-GFAP (rabbit polyclonal antibody; 1:1000; Cat#ab7260, Abcam, MA) for astrocytes, anti-GalC (mouse monoclonal antibody; 1:100; Cat#MAB342, Merck Millipore) for oligodendrocytes, and anti-A2B5 (mouse monoclonal antibody; 1:100; Cat#MAB312, Merck Millipore) for OPCs. The secondary antibodies used were Dylight 488-conjugated goat anti-rabbit IgG (1:200; Cat#E032220-2, EarthOx) and Alexa Fluor 594-conjugated goat anti-mouse IgG (1:200; Cat#A-11005, Invitrogen).