Anti stat1 10144 2 ap

The adipose tissues or RAW264.7 macrophages were lysed in RIPA Lysis Buffer in the presence of protease inhibitors. Cell lysates were centrifuged at 16,300 g for 10 min at 4°C. Subsequently, the Protein G agarose beads (Thermo Fisher) were conjugated with the anti-STAT1 (10144-2-AP, Proteintech) overnight at 4°C, followed by incubation with cell lysates overnight at 4°C with rotation. After washing, the immunoprecipitated proteins were eluted and subjected to Western blotting with the indicated antibodies.

The following primary antibodies were used in this study: mouse monoclonal anti-FLAG (M20008; Abmart), mouse monoclonal anti-β-actin (66009-1-lg; Proteintech), rabbit polyclonal anti-TMEM53 (HPA021134; Sigma-Aldrich), mouse monoclonal anti-dsRNA J2 (J2-1702; Scicons), rabbit polyclonal anti-STAT1 (10144-2-AP; Proteintech), mouse monoclonal anti-HA (TA180128; OriGene), rabbit polyclonal anti-HA (H6908; Sigma-Aldrich), and mouse anti-IgG (A7028, Beyotime). Anti-S-tag and anti-NP monoclonal antibodies were prepared in-house. We used the following secondary antibodies: horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (SA00001-1, Proteintech), HRP-conjugated goat anti-rabbit IgG (SA00001-2, Proteintech), goat anti-mouse IgG H&L DyLight 488 (ab96879, Abcam), goat anti-rabbit IgG H&L Cy3 (ab6939, Abcam), and goat anti-mouse IgG H&L Cy3 (SA00009-1, Proteintech). Universal type I interferon (IFN) (11200, PBL) was acquired from PBL Assay Science.