Recombinant Mouse Anti-VACV D13 Antibody (AD4) (CAT#: FAMAB-0348CQ)

Figure 1 Screen for murine hybridomas that secret anti-VACV antibodies with an immunofluorescence assay

HeLa cells grown on cover-slips were infected with VACV WR at a MOI of 0.1 to 0.5 PFU/cell for 8 hrs and then analyzed by immunofluorescence with hybridoma culture supernatants as the primary antibodies. The primary antibodies were stained with a Cy3-conjugated goat anti-mouse secondary antibody (red), and the DNA was stained with DAPI (blue). Specificity of the antibody to VACV is indicated by staining of only WR-infected cells, which display viral DNA factories as areas of cytoplasmic DNA staining. Images obtained with 11 representative hybridomas are shown with the hybridoma clone name. Shown in parenthesis is the group that the hybridoma was subsequently categorized into. Arrows point to viral factories (VF), virion-size particles (V) and aggregates (A).

Meng, X., Zhong, Y., Embry, A., Yan, B., Lu, S., Zhong, G., & Xiang, Y. (2011). Generation and characterization of a large panel of murine monoclonal antibodies against vaccinia virus. Virology, 409(2), 271-279.

Figure 2 Categorization of the hybridomas into 11 groups according to the apparent molecular weight of the precipitated protein

HeLa cells were infected with VACV WR at a MOI of 10 and metabolically labeled with 35S-methionine and -cysteine from 8 to 16 hpi. The cells were lyzed and immunoprecipitated with the hybridoma supernatants and Protein G sepharose. The precipitated proteins were analyzed by SDS-PAGE, and the autoradiographs of representative samples are shown. The results are grouped together according to the size of the precipitated proteins and are shown from the smallest to the largest precipitated proteins. Shown above each lane is the clone name of the hybrdioma whose supernatant was used for the immunoprecipiation. The control immunoprecipitation with DMEM-10 medium is indicated by "–". The group number of the hybridomas is indicated below the lanes.

Meng, X., Zhong, Y., Embry, A., Yan, B., Lu, S., Zhong, G., & Xiang, Y. (2011). Generation and characterization of a large panel of murine monoclonal antibodies against vaccinia virus. Virology, 409(2), 271-279.