Pierce co immunoprecipitation co ip kit

The interaction between BACH2 and FUS was examined in vivo using a Pierce Co‐Immunoprecipitation (Co‐IP) Kit (Thermo Fisher Scientific), according to the manufacturer's protocols. Coupling resin was incubated at 4 °C overnight with the indicated amounts of antibody. The antibody‐coupling resin complexes were then used to precipitate the cell lysates. Anti‐BACH2 (Cell Signaling Technology) and anti‐FUS (ProteinTech) were used to detect the precipitate. For in vitro binding assays, GSH‐agarose beads (Thermo Fisher Scientific) were used to purify the GST or GST‐BACH2 fusion bait protein, and His‐tag purification resin beads (Beyotime Biotechnology) were used to purify the His‐FUS fusion protein. GST protein or GST‐BACH2 fusion protein, which was combined with GSH‐agarose beads, was incubated with His‐FUS fusion protein for 6 h at 4 °C. The resulting bead‒protein‒protein complex was precipitated. Proteins isolated using elution buffer were detected by western blotting using anti‐GST (ProteinTech) and anti‐FUS (ProteinTech).

Anti-Cyr61, Anti-ITGAV, Anti-ITGB1, and goat Anti-Rabbit lgG H&L were from Abcam. Human recombinant Cyr61 was obtained from GeneTex. SiRNA-Cyr61 and negative control siRNA were purchased from Sangon Biotech (Shanghai). Pierce Co-Immunoprecipitation (Co-IP) Kit (26149) was purchased from Thermo scientific. The recombinant plasmid of pcDNA3.1-Cyr61 was constructed by our lab.