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. 2017 Jan 10;7:677. doi: 10.3389/fimmu.2016.00677

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Copyright © 2017 Kumar, Sunagar, Pham, Franz, Rosa, Hazlett and Gosselin.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Inactivated Francisella tularensis (iFt)-Mueller Hinton Broth (MHB) elicits antibody (Ab) against broader range of antigens compared to iFt-Brain-Heart Infusion (BHI). Ten micrograms of Ft LVS or WbtA LVS grown as indicated (M = MHB, B = BHI) were resolved by sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes for western blot analysis. Membranes were incubated overnight with 1:500 dilutions of the indicated sera. Bound Abs were detected through sequential incubations with biotinylated goat anti-mouse and streptavidin-conjugated horseradish peroxidase (HRP). The prominent ~18 kDa band visible in all three blots is an unidentified, endogenously biotinylated Ft protein detected by the SA-HRP conjugate.