polyclonal goat anti irf3 antibody  (R&D Systems)

Journal: PloS one

Article Title: TLR-9 contributes to the antiviral innate immune sensing of rodent parvoviruses MVMp and H-1PV by normal human immune cells.

doi: 10.1371/journal.pone.0055086

Figure Lengend Snippet: Figure 4. Defective replication of parvoviruses in hPBMCs. (A) hPBMCs collected from the blood of healthy donors were distributed into 6- well plates at 16107 cells/5 ml culture medium/well. They were then mock-treated or infected with MVMp or H-1PV at 10 PFUs/cell and 24 hrs later harvested for Southern blotting as described in Figure 2. The presented blot is representative of 3 additional which gave similar results. (B) hPBMCs (16107 cells/5 ml culture medium/well of a 6-well plate) and HEK293 (1.56106 cells/10-cm dish) were mock-treated or infected with the indicated parvovirus at 10 and 2 PFUs/cell respectively, and for the indicated period of time. Cultures were then harvested for Southern blotting as described in Figure 2. The presented blot is representative of 2 additional which gave similar results. (C) Expression of the parvovirus proteins in infected hPBMCs was examined in the same samples as those already analyzed for parvovirus replication in (B). At the time point indicated mock-treated or parvovirus- infected cultures were harvested by scraping in PBS and centrifuged. Cell pellets were then re-suspended in complete Ripa buffer supplemented with phosphatase and protease inhibitors. Total proteins were extracted from each sample as described in Materials and Methods. Fifty mg total proteins per sample were then subjected to 8% SDS-PAGE, transferred onto membranes, and probed with laboratory produced polyclonal antibodies specific for parvovirus NS1, NS2 and capsid polypeptides (Note: The antibody specific to VP proteins was designed to detect H-1PV polypeptides and is therefore less specific for MVMp). Actin was used as an internal loading control. The presented blot is representative of 2 additional which gave similar results. doi:10.1371/journal.pone.0055086.g004

Article Snippet: The mouse monoclonal anti-STAT1 and anti-PKR, as well as the rabbit polyclonal anti-STAT2 antibody and the goat polyclonal anti-IRF3 antibody were all from Santa Cruz Biotechnology (Heidelberg, Germany).

Techniques: Infection, Southern Blot, Expressing, SDS Page, Produced, Control

Journal: Journal of Virology

Article Title: Variation in Antagonism of the Interferon Response to Rotavirus NSP1 Results in Differential Infectivity in Mouse Embryonic Fibroblasts

doi: 10.1128/jvi.00585-09

Figure Lengend Snippet: FIG. 3. IRF3 nuclear translocation in wild-type MEFs following infection with the indicated strains of rotavirus. Wild-type MEFs were infected with the UK (A and B), RRV (C and D), ETD (E and F), UK-like reassortant 4-1-1 (G and H), or RRV-like reassortant 24-1-1 (I and J) strain, and IFNR KO MEFs were infected with UK (K and L) at an MOI of 0.5 for 6 h. The cells were stained for IRF3 (green), rotavirus VP6 (red), and nuclei (blue). The panels on the left show all three colors, except panel K, which shows only red and green, and the panels on the right show only IRF3 (green). The small arrows indicate antigen-positive cells with or without IRF3 nuclear translocation. The large arrows indicate antigen-negative cells with IRF3 nuclear trans- location. Magnification, 600.

Article Snippet: IRF3 was detected with a polyclonal goat anti-IRF3 antibody (R&D Systems, Inc.) and Alexa 488-labeled rabbit anti-goat immunoglobulin G (Invitrogen, Carlsbad, CA).

Techniques: Translocation Assay, Infection, Staining

Journal: Journal of Virology

Article Title: Variation in Antagonism of the Interferon Response to Rotavirus NSP1 Results in Differential Infectivity in Mouse Embryonic Fibroblasts

doi: 10.1128/jvi.00585-09

Figure Lengend Snippet: FIG. 4. Immune blot analysis of total IRF3 and pS396 IRF3 in wild-type MEFs infected with the indicated strains of rotavirus. Wild- type MEFs were infected with selected rotaviruses at an MOI of 30 for 16 h or mock infected. Proteins in the cell lysates were analyzed for total IRF3, pS396 IRF3 (pIRF3), and -actin by immune blotting.

Article Snippet: IRF3 was detected with a polyclonal goat anti-IRF3 antibody (R&D Systems, Inc.) and Alexa 488-labeled rabbit anti-goat immunoglobulin G (Invitrogen, Carlsbad, CA).

Techniques: Infection