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Western blot analysis of serum from chinchillas inoculated with the WT M. catarrhalis strain O35E.

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posted on 02.07.2013 by Teresa L. Shaffer, Rachel Balder, Sean W. Buskirk, Robert J. Hogan, Eric R. Lafontaine

Equivalent amounts of whole cell lysates (WT M. catarrhalis O35E, uspA2 KO strain O35E.2, hag transposon mutant strain O35E.TN2, and ompCD KO strain O35E.CD1) were resolved by SDS-PAGE, transferred to PVDF and probed with the indicated primary Abs. Panels A and B: Pre- and post-infection serum samples were pooled and used as primary Abs at a dilution of 1∶250. Goat α-rat IgG conjugated to HRP were used as secondary Abs. Controls: The murine monoclonal Abs 10F3 (Panel C, α-CopB), 5D2 (Panel D, α-Hag), 17H4 (Panel E, α-UspA2) and 1D3 (Panel F, α-OMPCD) were used as primary Abs in combination with goat α-mouse HRP-(IgG+IgA+IgM) secondary Abs. These controls were included to verify the identity of proteins recognized by post-infection chinchilla serum in panel B. MW markers are shown to the left of in kDa.