Mouse beta actin antibody

Western blot was performed according to the protocol described by Yang et al.^{31 (link)}. Briefly, whole-cell lysates were separated by SDS-PAGE, electro-transferred to PVDF membranes (Millipore), blocked for 1 h with 5% skim milk in TBS-T buffer (20 mM Tris-HCl [pH 7.4], 150 mM NaCl, and 0.1% Tween-20), and then incubated with the porcine circovirus type 2/PCV2 Capsid antibody (1:500, GeneTex, Texas, USA) or mouse Beta actin Antibody (1:2000, Proteintech Group, USA) at 4 °C overnight. Subsequently, the PVDF membranes were washed with 5% skim milk in TBS-T buffer three times, followed by blotting for 1.5 h with HRP-conjugated goat anti-rabbit IgG or HRP-conjugated goat anti-mouse IgG (1:5000, Beyotime, Jiangsu, China). The immunoreactive bands were visualized using the BeyoECL Plus Western blot detection system (Beyotime, Jiangsu, China) according to the manufacturer’s instructions. The experiments were repeated at least three times.

Luteolin and DTT were obtained from Aladdin (China). 1,4-butanediol diacrylate (BDD) was purchased from TCI (Shanghai, China). Dextran sulfate sodium (DSS, 35000 ​Da) was purchased from MP Biomedical (USA). Mouse Th1/Th2/Th17 CBA Kit, Cytofix/CytopermSoln Kit, Leuko Act Cktl with GolgiPlug, Transcription Factor Buffer Set, Fixable Viability Stain 780, FITC-Anti-CD4 antibody, BV605-Anti-IL-17A antibody, APC-Anti–INF–γ antibody and PE-Anti-IL-4 antibody were purchased from BD Biosciences (San Diego, USA). PE-Anti-FOXP3 antibody was purchased from eBiosciences (San Diego, CA). Myeloperoxidase (MPO) kit, glutathione (GSH) kit and ROS kit were supplied by Nanjing Jiancheng Bioengineering Institute (Nanjing, China). The BCA protein determination kit and Reactive Oxygen Species Assay Kit were purchased from Beyotime Biotechnology (China). Mouse Beta Actin antibody, Mouse Occludin antibody and Mouse ZO-1 antibody for western blotting were purchased from Proteintech (China). SYBR Premix Ex Taq™, Trizol reagent and PrimeScript™ RT 131 Master Mix were obtained from TaKaRa Bio (China). All other reagents are commercially available and can be used directly. All the solvents used were of analytical grade and were procured from Sinopharm (China).

SZ163 and SZ168, two mouse anti-hPDPN mAbs, were developed as described previously [24 (link)]. A mouse anti-hPDPN mAb (18H5), a normal mouse IgG (ab188776), and a rabbit anti-hPDPN mAb (EPR7072) were purchased from Abcam (Cambridge, UK). A mouse beta-actin antibody was purchased from ProteinTech (Wuhan, China). Fluorescein isothiocyanate-conjugated goat anti-mouse IgG polyclonal antibodies (FITC-GAM IgG) were purchased from Beckman-Coulter (Suzhou, China).