Anti jak2 antibody

Total proteins in ASMCs were extracted with RIPA lysis buffer (Solarbio, Beijing, China),
and their concentration was quantified by a BCA protein detection kit (Solarbio, Beijing,
China). An equal amount of protein samples (20 μg per group) was separated by SDS-PAGE and
transferred to a polyvinylidene fluoride membrane (Millipore, Bedford, MA, USA), which was
then blocked with 5% skimmed milk for 1 hour. The membrane was firstly incubated with the
primary antibody at 4°C overnight, and then with the secondary antibody, goat anti-rabbit
IgG H&L (Abcam, ab6721, 1: 3000) at 37°C for 1 hour. Ultimately, the bands were
developed with the enhanced chemiluminescence reagent (Pierce Biotechnology, Rockford, IL,
USA). The primary antibodies were: anti-HMGB1 (1: 1000, ab79823),
anti-p-JAK2 (1: 1000, #8082), anti-p-STAT3 (1: 1000,
#9145), anti-JKA2 (1: 1000, #3230), anti-STAT3 (1: 1000,
#12640) and anti-β-actin (1: 1000, ab6276). Among them,
anti-HMGB1 antibody and antiβ-actin antibody were
bought from Abcam (Shanghai, China); anti-p-JAK2 antibody,
anti-p-STAT3 antibody, anti-JAK2 antibody, and
anti-STAT3 antibody were all purchased from Cell Signaling Technology
(Cell Signaling Technology, Danvers, MA, USA).