Goat anti mouse igg conjugated to fluorescein isothiocyanate fitc

Primary antibodies used for immunofluorescence assays were: mouse mAb anti-glycoprotein E of DENV (ab41349, Abcam), mouse mAb SA02-BG12 anti N protein of JUNV (Sánchez et al., 1989) , mouse mAb anti-hnRNP A2 (ab6102, Abcam), rabbit Ab anti-hnRNP K (R332, Cell Signaling), rabbit Ab anti-T7 (ab9115, Abcam) and rabbit immunosera obtained in our laboratory: anti-DENV-2, anti-JUNV and anti-VSV. Goat anti-mouse IgG conjugated to fluorescein isothiocyanate (FITC) or rhodamine (TRITC), used as secondary antibodies were purchased from Sigma-Aldrich.
Western blot analysis was performed employing mouse mAb anti-hnRNP A2 (ab6102, Abcam), rabbit Ab anti hnRNP K (R332, Cell Signalling), mouse mAb anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH, ab8245, Abcam), rabbit Ab anti-T7 (ab9115, Abcam) or rabbit Ab anti-histone H4 (sc-8660 R, Santa Cruz Biotechnology) as primary antibodies and peroxidase anti-rabbit IgG (W4011, Promega) or peroxidase anti-mouse Ig G (A9044, Sigma-Aldrich) as secondary antibodies.

The specificity of the antibody response elicited by immunization was tested by immunofluorescence assays (IFA) using airdried thin films of erythrocytes infected with P. vivax schizonts as described previously [39] . Surface expression was detected using sera from mice immunized with synthetic peptides RII, pLRII and TTRII and affinity-purified goat anti-mouse IgG conjugated to fluorescein isothiocyanate (FITC) (Sigma, St. Louis). Pools of sera from animals of each group (RII, TTRII, pLRII) collected on days 63 and 84 were tested at 1:50 dilution. DAPI (4 0 ,6-Diamidine-2 0 -p henylindole dihydrochloride) (SIGMA, St. Loius) was used to confirm the presence of DNA. Serum of an individual from Brazilian Amazon, who presented high antibody titers against P. vivax merozoite proteins was used as positive control.