|Detection of Human IRF3 by Western Blot. Western blot shows lysates of Raji human Burkitt's lymphoma cell line, Daudi human Burkitt's lymphoma cell line, and Jurkat human acute T cell leukemia cell line. PVDF Membrane was probed with 1 µg/mL of Mouse Anti-Human IRF3 Monoclonal Antibody (Catalog # MAB4019) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for IRF3 at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
Intracellular Staining by Flow Cytometry
|Detection of IRF3 in Daudi Human Cell Line by Flow Cytometry. Daudi human Burkitt's lymphoma cell line was stained with Mouse Anti-Human IRF3 Monoclonal Antibody (Catalog # MAB4019, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005).View our protocol for Staining Intracellular Molecules.|
IRF3 (interferon response factor 3) is a 60 kDa member of the IRF family of proteins. Human IRF3 contains one DNA binding domain (aa 7‑107), a nuclear export signal (aa 139‑149) and multiple phosphorylation sites (aa 395‑407). Viral infection stimulates IRF3 phosphorylation, nuclear translocation and stimulation of IFN production. Alternate splice forms may exist. One will show a deletion of aa 201‑327, a second will show the same deletion plus an alternate start site at Met147, and a third will show a 125 aa substitution for the C-terminal 100 aa (aa 328‑427). Over aa 206‑427, human IRF3 is 76% and 83% aa identical to mouse and pig IRF3, respectively.