Surface protein fraction was separated by 2-DE and probed with mouse anti- C. perfringens (heat killed whole cell) serum. Goat anti-mouse HRP conjugate was used as secondary antibody (1:2000 dilutions) and blots were developed using Immuno-Blot HRP assay kit (Bio-Rad, USA) as per manufacturer’s instructions. A, Coomassie stained 2-DE gel; B, corresponding

blot as described above. Spots identified in this study are indicated with arrows. (TIFF 1 MB) Additional file 6: Proteins identified in this study and their homologues BI 10773 nmr in other bacteria. A few pathogenic organisms where the presence of respective protein has been shown experimentally in other studies are listed along with their localization and predicted role. (DOC 165 KB) Additional file 7: Pattern/profile, post translational modifications and topology search results for identified proteins of Clostridium perfringens. Proteins identified from different fractions, indicating theoretical localization. All the analysis was carried out using ExPASy Proteomics tools at http://​www.​expasy.​ch. (DOC 104 KB) References 1. MacLennan JD: Anaerobic infections of war wounds in the Middle East. Lancet 1943, ii:123–126.CrossRef 2. Rood IR, Cole ST: Molecular genetics and pathogenesis of Clostridium perfringens. Microbiol Rev 1991, 55:621–648.PubMed 3.

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with C-domain of α-toxin prevents lethal infection, localizes tissue injury, and promotes host response to challenge with Clostridium perfringens. J Infect Dis 2004, 190:767–773.CrossRefPubMed 7. Titball RW, Naylor CE, Moss D, Williamson ED, Basak AK: Mechanism of protection against disease caused by Clostridium perfringens. Immunology 1998, 95:34. 8. Calabi E, Fairweather N: Patterns of sequence conservation in the S-layer proteins and related sequences in Clostridium difficile. J Bacteriol 2002, 184:3886–3897.CrossRefPubMed 9. DelVecchio VG, Connolly JP, Alefantis TG, Walz A, Quan MA, Patra G, Ashton JM, Whittington JT, Chafin RD, Liang X, Grewal P, Khan AS, Mujer CV: Proteomic profiling and identification of immunodominant spore antigens of Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis. Appl Env Microbiol 2006, 72:6355–6363.CrossRef 10.