Anti bace2 antibody

Cells were lysed with RIPA-Doc buffer (Tris-HCl, 50 mM; NaCl, 150 mM; Triton X-100, 1%; deoxycholate, 1%; and SDS, 0.1%; supplemented with 1/100 protease inhibitors). Cell lysates were separated by 10% Tris-glycine SDS-PAGE gels and transferred to PVDF membranes. The membranes were blocked in 5% non-fat milk for 1 h, then incubated overnight at 4 °C with anti-myc (9E10), anti-BACE2 and anti-β-actin (AC-15) antibodies, respectively [18 (link)]. The membranes were washed in TBST with 0.1% Tween-20 and incubated with HRP-labeled goat anti-mouse or HRP-labelled goat anti-rabbit antibodies at room temperature for 1 h. Anti-myc antibody 9E10 was obtained from Abcam. Anti-BACE2 antibody was purchased from Santa Cruz (sc-271,212). β-actin antibody AC-15, HRP-labelled goat anti-rabbit antibody and HRP-labelled goat anti-mouse antibody were obtained from ZSGB-BIO. The image was obtained by using FluorChem R imaging system.

Inhibitors LY2886721, Lanabecestat, and Marimastat were from Selleck Chemicals and TMI-1 was from Tocris Sciences. Anti-ADAM17 and anti-ADAM9 were from Cell Signaling Technology (Cat:3976S and 4151S, 1:000 dilution), anti-BACE2 antibody was from Santa Cruz Biotechnology (sc-271212, 1:1000 dilution), and anti-MSLN antibody (mouse monoclonal MN, 1 ug/ml) was from our lab. siRNAs against human ADAM17: s13719 (siADAM17_1) and si13720 (siADAM17_2) and one siRNA targeting ADAM10: s1004 were from Thermo Fisher Scientific. All other On-Target Plus siRNA and negative control siRNA luciferase GL2 were from Dharmacon.