Anti zikv e antibody

RIPA buffer (Sigma-Aldrich) plus a protease and phosphatase inhibitor cocktail (Roche, Basel, Switzerland) was used to lyse cells as previously described [18 (link)]. Proteins were loaded onto SDS-PAGE gels, transferred onto polyvinylidene difluoride membranes, and blocked with 5% skim milk in Tris-buffered saline supplemented with 0.1% Tween-20 (TBS-T) for 1 h at room temperature. The membranes were incubated overnight with primary antibodies at 4 °C. The following antibodies were used; Anti-FLAG tag (Wako), Anti-MYC tag (Abcam), Anti-GFP (Santa Cruz), Anti-Tubulin or β-actin antibodies (Abgent). Anti-pSTAT1/STAT1, Anti-HA tag, Anti-HIS tag, Anti-MYC tag, Anti-MDA5, Anti-RIG-I, Anti-pTBK1/TBK1, Anti-pIRF3/IRF3, Anti-MxA, Anti-OAS1, and Anti-RNaseL antibodies were from Cell Signaling Technology. Anti-SARS-CoV/SARS-CoV-2 Nucleocapsid Antibody (Sino Biological), while Anti-IFV NP antibody and Anti-ZIKV E antibody were purchased from GeneTex. The blots were washed with TBST for three times and incubated with HRP-conjugated secondary antibodies (Cell Signaling Technology, Danvers, MA, USA). Proteins were visualized with ECL Western Blotting Substrate (Thermo Fisher Scientific, Waltham, MA, USA). Densitometry to quantify immunoblot bands was performed using ImageJ software.

Restriction enzymes, DNA-modifying enzymes, the Q5 High Fidelity PCR kit and the ProtoScript II First Strand cDNA synthesis kit were obtained from New England Biolabs (Ipswich, MA, USA). SuperScript II was obtained from Invitrogen (Carlsbad, CA, USA).
Anti-flavivirus monoclonal antibody D1-4G2-4-15, which reacts with ZIKV E protein, was obtained from EMD Millipore (Billerica, MA, USA). Secondary antibodies were obtained from Sigma (St. Louis, MO, USA) and Invitrogen. Oligonucleotide primers and probes for DNA amplification and quantitative PCR (qPCR) were obtained from Sigma and IDT (Coralville, IA, USA). Anti-ZIKV E antibody was obtained from Genetex (Cat # GTX1333325). Anti-ZIKV E DIII monoclonal antibody (ZV-57) was a kind gift from Michael Diamond, Washington University School of Medicine, St. Louis, MO, USA.