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. Author manuscript; available in PMC: 2011 Sep 15.
Published in final edited form as: Virology. 2010 Jun 17;405(1):41–49. doi: 10.1016/j.virol.2010.05.025

Fig. 6.

Fig. 6

Localization of VACV neutralizing epitope on the A28 protein. (A) Diagram of A28 truncations. Representation of A28 protein with amino acid numbers is shown at the top. A28T-1 shows the A28 protein with replacement of the 26-amino acid N-terminal transmembrane domain by the Ig μ-chain signal peptide sequence (SS) and addition of a six-histidine (6xHis) tag at the C-terminus. A28T-1 to A28T-4 constructs contain serial N-terminal truncations but retain the signal peptide and six-histidine tag. A28T-5 to A28-T7 constructs have serial C-terminal truncations and A28T-8 has both N- and C-terminal truncations. (B) Western blot. Plasmids encoding truncated proteins were transfected into HeLa cells and the lysates were analyzed by Western blotting with antibody to the histidine tag. (C) The A28 proteins diagrammed in panel A were expressed by transfection and bound to beads via the six-histidine tag. The beads were incubated with A28/H2 sera and the depleted sera were tested for neutralizing activity. A representative of three assays is shown.