Rabbit anti myc

Processing of transfected cells for confocal microscopy was performed as we described previously. Briefly, monolayers were permeabilized and fixed using 100% methanol held at –20°C for 5 min and were then blocked and labeled with mouse-anti-FLAG (Sigma) (1/200 dilution in 3% bovine serum albumin [BSA]–phosphate-buffered saline [PBS]) and rabbit-anti-Myc (Proteintech) (1/200 dilution in 3% BSA–PBS). Cells were counterlabeled with Alexa Fluor 488 anti-mouse antibody (Invitrogen) (1/4,000 dilution in 3% BSA–PBS), Alexa-Fluor 555 anti-rabbit antibody (Invitrogen) (1/4,000 dilution), and DAPI (4′,6-diamidino-2-phenylindole) to stain the nuclei. Monolayers were examined by confocal microscopy. A total of 100 cells for each replicate were counted to determine the presence or absence of localization.

The following antibodies were used: rabbit anti-DYKDDDDK tag (ABclonal, Cat # AE005), rabbit anti-HA (CST, Cat # 3724S), rabbit anti-IRF3 (ABclonal, Cat # D199862-0100), rabbit anti-p-IRF3 (CST, Cat # 4947S), rabbit anti-TBK1 (4A Biotech co.Ltd, Cat# 4ab032308cs), rabbit anti-p-TBK1 (Absin, Cat # abs140019), rabbit anti-KPNA6 (ImmunoWay, Cat # YN3049), rabbit anti-STAT2 (CST, Cat # 72604S), rabbit anti-p-STAT2 (CST, Cat # 88410S), rabbit anti-p-STAT1 (CST, Cat # 9167S), rabbit anti-p-TYK2 (Absin, Cat # abs131318), rabbit anti-TYK2 (Absin, Cat # abs131318a), rabbit anti-Myc (Proteintech, Cat # I6286-I-AP), anti-rabbit IgG HRP-linked antibody (CST, Cat # 7074), mouse anti-DYKDDDDK tag (Bioworld, Cat # AP0007), mouse anti-GAPDH (Proteintech, Cat # 60004-I-Ig), mouse anti-VSV-G (Abgent, Cat # AP1016a) and HRP goat anti-mouse IgG (BioLegend, Cat # 405306). Alexa Fluor 488 goat anti-mouse IgG (Thermo Fisher Scientific) and Alexa Fluor 647 goat anti-rabbit IgG secondary antibodies (Thermo Fisher Scientific) were employed, as well as protein A/G beads (Bimake) and anti-Flag magnetic beads (Bimake).
The following reagents were used: transfection reagent and PEI power (Aladdin, USA), Protease inhibitor (Sigma-Aldrich, USA), and recombinant human IFN-α (PeproTech, USA).