Hrp conjugated donkey anti goat igg sc 2020

Cells were seeded into 12-well plates for 24 h and then treated with various concentrations of human IFN-β1a or mouse IFN-β (PBL) as indicated in each experiment. Cells were infected with Candid#1 JUNV at an MOI of 3 PFU/cell. IFNs were supplemented after virus infection. Protein lysates were prepared in 2x Laemmli sample buffer at 1 and 2 days p.i. from MEF cells and A549 cells, or from Vero cells at 2 days p.i.. Protein samples were resolved on 4–20% SDS-PAGE gel and transferred to PVDF membranes using Mini Trans-Blot Electrophoretic Transfer Cell apparatus (Bio-Rad, CA). Membranes were incubated with primary antibodies overnight at 4°C and then with appropriate secondary antibodies for 1 h at room temperature. Proteins were visualized with ECL Western Blotting Detection Reagents (GE, NJ) according to the manufacturer's instruction. Viral NP protein was detected with a monoclonal mouse anti-JUNV NP antibody (AG12, BEI). Equal loading of samples was confirmed by immunoblotting of the same membranes with an antibody to β-actin (sc-1616, Santa Cruz). Secondary antibodies HRP-conjugated Goat anti-mouse IgG (115-035-146, Jackson Immunology) and HRP-conjugated donkey anti-goat IgG (sc-2020, Santa Cruz) were used.

Mouse monoclonal anti-GAPDH antibody (sc-32233), rabbit polyclonal anti-ERK1 antibody sc-94), mouse monoclonal anti-pERK antibody (sc-7383), and mouse monoclonal anti-histone H1 antibody (sc-8030) were purchased from Santa Cruz Biotechnology (Dallas, TX) and used for Western blot (WB) analysis. Anti-PGC-1α antibody (ab54481) was obtained from Abcam (Cambridge, UK) and used for WB and IHC analyses. Anti-8-OHdG antibody (MOG-020P) was obtained from the Japan Institute for the Control of Aging (Shizuoka, Japan). Rabbit polyclonal anti-p38 MAPK antibody and rabbit polyclonal anti-phospho-p38 MAPK were obtained from Cell Signaling Technology, Japan (Tokyo, Japan). Horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (170-6515; Bio-Rad) and HRP-conjugated donkey anti-goat IgG (sc-2020; Santa Cruz Biotechnology) were used as secondary antibodies for WB analysis.

The anti-MRTF-A/B and anti-caldesmon antibodies were described previously [16 (link), 45 (link)]. Anti-tropomyosin (T2080), anti-α-actin (A5441), anti-vinculin (V9131), anti-Talin (T3287), anti-tubulin (T9024), and anti-FLAG (F7425) antibodies were from Sigma. Anti-SRF (sc-335), anti-zyxin (sc-6437), anti-Myc (sc-789), anti-RhoA (sc-418), anti-MYPT1 (sc-25618) and anti-c-Src (sc-18) antibodies were from Santa Cruz. Anti-pY118-phosphopaxillin antibody (#3818-1) was from Epitomics, anti-paxillin (MA3060) and anti-integrin αv (AB1930) antibodies from Chemicon, anti-GAPDH antibody (M171-3) from MBL, anti-HA antibody (3F10) from Roche, anti-integrin β1 antibody (610467) from BD Transduction Laboratories, and anti-FLAG antibody (018-22386) from Wako. Anti-FAK (#3285), anti-pY397-phosphoFAK (#3283), anti-pY576/577-phosphoFAK (#3281), anti-pY925-phosphoFAK (#3284), anti-pY527-phosphoSrc (#2105), anti-pY416-phosphoSrc family (#6943) and anti-pT696-phosphoMYPT1 (#5163) antibodies were from Cell Signaling Technology. The secondary antibodies, horseradish peroxidase (HRP)-conjugated donkey anti-rabbit IgG (NA934) and sheep anti-mouse IgG (NA931) were from GE Healthcare, HRP-conjugated donkey anti-goat IgG (sc-2020) from Santa Cruz, and Alexa-conjugated goat anti-rabbit IgG (A-11034, A-11036) and goat anti-mouse (A-21049, A-11031) from Thermo Fisher Scientific.