Goat anti rabbit igg h l hrp

The primary antibodies used in this study were at the dilutions of 1:1000 as follows: CD229 (ab103172, Abcam, UK); RASAL3 (ARP79758_P050, Aviva Systems Biology, USA); Flag (14793S, Cell Signaling Technology, USA); ERK (4695S, Cell Signaling Technology, USA); p-ERK (4370S, Cell Signaling Technology, USA); GAPDH (5174S, Cell Signaling Technology, USA). The second antibodies Goat anti-Rabbit IgG (H+L) HRP (FMS-Rb01, Fcmacs) and Goat anti-Mouse IgG (H+L) HRP (S0002, Affinity) were in 5000 diluted concentrations. Puromycin was obtained from Merck KGaA (Darmstadt, Germany). Diphenyltetrazolium Bromide (MTT) was purchased from Solarbio (Shanghai, China). PKH26 was purchased from Sigma-Aldrich (Lot#SLBP9768V, SIGMA, USA).

Reagents are as follows: purified water (Wahaha Group); acetonitrile (chromatographic grade, Fisher Scientific); methanol (chromatographic grade, Yuwang Industrial Co., Ltd.); IL-4 (JL21520), IFN-γ (JL11665), IgE (JL11287) ELISA kits (Shanghai Jianglai Biotechnology Co., Ltd.); FLG (Affinity, USA, DF13653); LOR (Thermo Fisher Scientific, USA); Caspase-14 (Novus, USA); 0.25% trypsin protein digestion solution (Beijing Soleibao Technology Co., Ltd., T1350); goat anti-rabbit secondary antibody, goat anti-mouse secondary antibody, DAB color reagent, ECL luminescence kit (Beijing Zhongshan Jinqiao Biotechnology Co., Ltd., ZB2301, ZB2305, ZLI-9018, SC-2048); hematoxylin staining solution (Zhuhai Besuo Biotechnology Co., Ltd., BA-4041). The second antibodies goat anti-rabbit IgG (H+L) HRP (FMS-Rb01, Fcmacs) and goat anti-mouse IgG (H+L) HRP (S0002, Affinity) were in 5000 diluted concentrations.

The following antibodies were used: anti-SRD5A1 (Proteintech, #66329-1-Ig, 1:1,000 dilution), anti-nuclear factor-κB (NF-κB) p65 (Cell Signaling Technology, #8242, 1:1,000 dilution), anti-Phospho-NF-κB p65 (Cell Signaling Technology, #3033, 1:1,000 dilution), anti-VEGF (Proteintech, #19003-1-AP, 1:1,000 dilution), and anti-β-actin (Proteintech, #66009-1-Ig, 1:5,000 dilution). Secondary antibodies included goat anti-Rabbit IgG(H+L) HRP (Fcmacs, #FMS-Rb01, 1:5,000 dilution) or mouse (Affinity, #S0002, 1:10,000 dilution). 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) was purchased from Solarbio (Beijing, China), dutasteride was purchased from APExBIO (Houston, TX, USA), propidium iodide (PI) was purchased from XunBei (Nanjing, China), and Annexin-V-FITC was purchased from Biolegend (San Diego, CA, USA). Pierce TM BCA Protein Assay kit, transfection reagent, and RNase A were purchased from YEASEN (Shanghai, China). Senescence-associated β-galactosidase (SA-β-gal) kit, puromycin, and doxycycline were purchased from Beyotime Institute of Biotechnology (Shanghai, China). TRIeasy™ Total RNA Extraction Reagent, complementary DNA synthesis superMix, and SYBR Green Master Mix were purchased from YEASEN (Shanghai, China).