Mouse anti ha tag mab

Cell lysates were prepared in RIPA buffer and subject to SDS-PAGE and immunoblotting analysis were described previously [25 (link), 43 (link)]. Immunofluorescence staining were performed as previously described [25 (link), 43 (link)]. The following monoclonal (mAb) and polyclonal (pAb) antibodies were utilized: mouse anti-Flag-tag M2 mAb (Sigma); mouse anti-HA-tag mAb (Invivogen); rabbit anti-maltose-binding protein (MBP) pAb (New England Biolabs); mouse anti-flavivirus envelope protein (4G2) mAb (Millipore); mouse anti-ZIKV NS5 mAb, clone 8B8 (Biofront Technologies, kindly provided by Hengli Tang, Florida State University); mouse anti-β-actin mAb (Sigma); rabbit anti-TRIM56 pAb (Bethyl Labs) or rabbit anti-TRIM56 S4091 pAb (generated by immunizing rabbits at Proteintech Group Inc. with a recombinant protein comprising the C-terminal 392 aa of human TRIM56 fused to MBP that was expressed and purified from E. coli); peroxidase-conjugated secondary goat anti-rabbit and goat anti-mouse pAbs (Southern Biotech); FITC-conjugated secondary goat anti-mouse pAb (Southern Biotech). Specifically, FH-T56 was detected by mouse anti-Flag-tag M2 mAb (Sigma); rabbit anti-TRIM56 S4091 pAb was used to detect endogenous T56 protein in HeLa cell lysates; and rabbit anti-TRIM56 pAb (Bethyl Labs) was used for other T56 immunoblotting experiments.

Cell lysates were prepared and subject to immunoprecipitation or/and immunoblot analysis as previously described (15 (link), 46 (link)). The following pAbs and mAbs were utilized: rabbit anti-TRIM56 S4091 pAb (this study); rabbit anti-pSer⁴⁷¹-TRIM56 and anti-pSer⁴⁷⁵ pAbs (this study); rabbit anti-ISG56 pAb (42 (link)); rabbit anti-TRIF pAb (51 (link)); rabbit anti-IRF-3 pAb (52 (link)); rabbit anti-pSer³⁹⁸-IRF3 (Upstate), rabbit anti-MBP pAb (New England Biolabs), mouse anti-TRIM56 mAbs (this study), mouse anti-HA tag mAb (Invivogen); mouse anti-V5 tag mAb (Invitrogen); mouse anti-Flag M2 mAb (Sigma); mouse anti-actin mAb and rabbit anti-calreticulin pAb (both form Sigma); peroxidase-conjugated secondary goat anti-rabbit and goat anti-mouse pAbs (Southern Biotech).