Goat anti rabbit igg h l

T2 was obtained from Sigma-Aldrich (St. Louis, MO, USA) and dissolved in DMEM at 10 µM. Primary antibodies (rabbit anti-β-actin pAb (CST, Danvers, MA, USA) and mouse anti-IκBα mAb (Proteintech, Rosemont, IL, USA)) and secondary antibodies applied for Western blotting (goat anti-mouse IgG (H+L) (Promega, Madison, WI, USA) and goat anti-rabbit IgG (H+L) (Promega, Madison, WI, USA)) were purchased from commercial companies. Cyanine 5(Cy5)-conjugated goat anti-mouse IgG used in confocal microscopy was obtained from Gibco BRL life Technologies (New York City, NY, USA). Mouse monoclonal anti-cleaved-caspase-3 antibody and anti-cleaved-caspase-8 were purchased from Servicebio (Hubei, Wuhan, China). Rabbit polyclonal anti-Nrf2 antibody, anti-Gpx-1, mouse monoclonal anti-Bax antibody, and anti-Bcl-2 were purchased from Bioss (Beijing, China). In addition, the primary antibody mouse anti-PRV-gB mAb was a gift obtained from Dr. Ping Jiang (College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China).

Cells were cultured in 6-well plates. The infected cells were collected after infection 48 and 72 h and lysed in 200 μL of Passive Lysis Buffer (Promega, USA), and the protein concentration was determined using a BCA protein quantification kit (Genesand, China). Equal amounts of protein samples were separated via SDS-PAGE and transferred to a 0.45 µm PVDF membrane (Millipore, USA). The protein expression levels of ISKNV were measured by a mouse anti-isknvORF108L (an ISKNV viral structural protein) antibody (mAb2D8, from Dr. Chuanfu Dong). The antibody was also named anti-ISKNV-VP101L. To manifest equal protein sample loading, S. chuatsi GAPDH expression was measured using rabbit anti-GAPDH (Abways, China). Goat anti-mouse IgG (H + L) and goat anti-rabbit IgG (H + L) (Promega, USA) were used as the secondary antibodies. Finally, the images were acquired from Amersham Image 600 using a High-sig ECL Western blotting substrate kit (Tanon, China).