Beclin 1 antibody

Ovalbumin (OVA) was obtained from Sigma (St.Louis, MO, USA). Rhynchophylline was obtained from Melonepharma (Dalian, China) or Aladdin (Shanghai, China). Transforming growth factor (TGF)-β1 was obtained from USCN (Wuhan, China). WP1066 was obtained from Aladdin. Mouse interleukin (IL)-6 enzyme-linked immunosorbent assay (ELISA) kit and IL-13 ELISA kit were obtained from BOSTER (Wuhan, China) and IgE ELISA kit was obtained from Multi Science (Hangzhou, China). Catalase (CAT) detection kit and superoxide dismutase (SOD) detection kit were obtained from JianchengBio (Nanjing,China). Primary antibodies against LC3, ATG5, p62, Janus kinase (JAK) 2, p-JAK2 and STAT3 were obtained from Abclonal (Wuhan, China). Beclin-1 antibody and β-actin antibody were obtained from Santa Cruz (Dallas, TX, USA). P-STAT3 antibody was obtained from Affinity (Changzhou, China). Histone H3 antibody was obtained from ABGENT (San Diego, CA, USA). α-SMA antibody was obtained from Abcam (Cambridge, UK). Calponin antibody was obtained from Proteintech (Wuhan, China). Horse radish peroxidase-labeled, Cy3-labelled and fluorescein isothiocyanate (FITC)-labeled secondary antibodies were obtained from Beyotime (Shanghai, China).

Protein samples were separated on a 1% agarose
gel under denaturing conditions and electrophoretically
transferred to nitrocellulose
membranes. The membranes were probed with
1:500 dilution of Beclin-1antibody (sc-11427,
Santa Cruz Biotechnology, Inc., Santa Cruz, CA,
U.S.) and then incubated with a secondary antibody
(anti-mouse IgG, 1:5000; Beyotime Biotech,
Shanghai, China). For detection, enhanced
chemiluminescence was carried out with BCA
kits (Beyotime Biotech, Shanghai, China). An
internal reference sample (the same one for each
blot) was included as a standard for quantification.
The signal from each band was measured
against the standard, and this relative number
was used for statistical analysis.