Anti gst

The mouse monoclonal anti-LRP6 antibody was from Santa Cruz (Santa Cruz, CA, USA). Rabbit polyclonal anti-p-LRP6 (Ser1490) was from Bioss (Woburn, MA, USA). Mouse monoclonal anti-β-catenin was from BD (Franklin Lakes, NJ, USA). Anti-phalloidin was from Life (Carlsbad, CA, USA). Mouse monoclonal anti-vinculin was from Boster (Wuhan, China). Mouse monoclonal anti-α-tubulin, acetyl-α-tubulin, tyrosine-α-tubulin were from Sigma-Aldrich (Saint Louis, MO, USA). Rabbit polyclonal anti-detyrosinated-α-tubulin, Mouse monoclonal anti-RhoA, Rac1and Cdc42 were purchased from Abcam (Cambridge, MA, USA). Rabbit polyclonal anti-phosphor-MAP1B (Thr1265) was from Novusbio (Littleton, CO, USA). Rabbit polyclonal anti-MACF1 was from Proteintech (Chicago, IL, USA). Rabbit monoclonal anti-phosphor-GSK3β (Ser9) and mouse monoclonal anti-GSK3β were from Cell Signaling (Beverly, MA, USA). Rabbit polyclonal anti-His, mouse monoclonal anti-His and anti-GST were from Origene (Rockville, MD, USA).

Anti-Flag (Sigma-Aldrich), anti-Myc (Santa Cruz), anti-α-tubulin (Sigma-Aldrich), anti-His (Origene), anti-GST (Origene), and anti-HA (Origene) mouse monoclonal antibodies as well as anti-IL-1β (Santa Cruz) rabbit polyclonal antibodies were used as primary antibodies for immunoblotting and immunoprecipitation. HRP-conjugated goat anti-rabbit (ZSGB-BIO) or anti-mouse IgG (ZSGB-BIO) antibodies were used as secondary antibodies for immunoblotting.
Immunoprecipitation was performed using IP buffer (1% Nonidet P-40, 50 mM Tris-HCl [pH 7.5], 150 mM NaCl, and Complete™ protease inhibitor cocktail-EDTA (Roche)). Whole cell extracts were prepared after transfection and incubated with indicated antibodies together with Protein A/G beads (Roche) overnight. Beads were then washed 4 times with IP buffer, and immunoprecipitates were eluted with SDS loading buffer (TransGen Biotech) and resolved in SDS-PAGE gels. The proteins were transferred to PVDF membrane (Bio-Rad) and further incubated with the indicated antibodies. The antigen-antibody complexes were visualized by the Immubilon™ chemiluminescent detection kit (Millipore).