Anti actin ab1801

Samples used for the WB experiments were adjusted to equal protein concentration in advance.
For the determination of BACE1, cell lysates were prepared by lysing cells in 150 mM NaCl, 50 mM Tris/HCl pH 7.4, 2 mM EDTA, 0.1% NP-40, 0.1% Triton-X 100. For the determination of total secreted Aβ level and sAPPβ conditioned media were used.
The following antibodies were used for WB analysis: W02 antibody (5 μg/mL; Millipore, Billerica, MA, USA), anti-sAPPβ: Mbs492139 (1:250; MyBioSource, San Diego, CA, USA), BACE1: ab2077 (1:1000; abcam, Cambridge, UK), anti-actin ab1801 (1:1000; abcam), anti-rabbit IgG HRP Conjugate W401B (1:5000; Promega, Mannheim, Germany) and anti-mouse P0260 (Dako, Hamburg, Germany).
Aβ levels were detected by performing immunoprecipitation of conditioned media before WB analysis. Therefore, 20 μL protein G-Sepharose and W02 antibody (5 μg/mL) were used.
Enhanced chemiluminescense (ECL)-method (Perkin Elmer, Rodgau-Jügesheim, Germany) was used to detect proteins. Densitometrically quantification was performed by using Image Gauge V3.45 software (Fujifilm, Düsseldorf, Germany).

DON was obtained from Sigma-Aldrich (St. Louis, MO, USA). PDTC was obtained from Beyotime (Shanghai, P.R. China). Anti-iNOS (ab15323) and anti-actin (ab1801) antibodies were purchased from Abcam (Cambridge, MA, USA). Anti-phospho-IκBα (Ser32/36; 5A5), anti-phosphoryl-NF-κB p65 (Ser536; 93H1), and peroxidase-coupled goat anti-rabbit and mouse IgG (H + L) secondary antibodies were obtained from CST (Danvers, MA, USA). The apoptosis detection kit (Annexin V-FITC) was purchased from BestBio (Shanghai, China).