Download E. coli. Shiga Toxin Methods and Protocols by Dana Philpott, Frank Ebel PDF
By Dana Philpott, Frank Ebel
A multidisciplinary panel of prime experimentalists information the major protocols for the analysis and learn of the pathogenesis of Shiga toxin-producing Escherichia coli (STEC) and its remoted cytotoxin (Stx). defined in step by step aspect, those with no trouble reproducible innovations diversity from these for the prognosis and detection of STEC micro organism in sufferer and animal samples, to these for learning the mobile microbiology of STEC infections, specially host-pathogen interactions and the hemolysin of STEC. There also are protocols for learning the main points of Shiga toxin (Stx) biology-from the purification of the toxin to the results of Stx on a variety of host telephone functions-and for exploring STEC-mediated ailment in a number of animal versions.
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Acknowledgments The authors thank Olga Böhler and Dr. Kerstin Ludwig and Dr. Martina Bielaszewska for sharing protocols and technical advice. References 1. Karmali, M. A. (1989) Infection by verocytotoxin-producing Escherichia coli. Clin. Microbiol. Rev. 2, 15–38. 2. Paton, J. C. and Paton, A. W. (1998) Pathogenesis and diagnosis of Shiga toxinproducing Escherichia coli infections. Clin. Microbiol. Rev. 11, 450–479. 3. Karmali, M. , Fleming, P. , Arbus, G. , and Lior, H. (1985) The association between idiopathic hemolytic uremic syndrome and infection by verotoxin-producing Escherichia coli.
Dry overnight at room temperature. 26F 33 8/1/2002, 3:44 PM 34 Bitzan and Karch Fig. 1. E. coli O111:B4 lipopolysaccharide (Sigma), 6 µg/lane, fractionated by denaturing SDS-PAGE and silver stained, with protein molecular-weight markers of the indicated sizes. tion of 10 or 20 µg/mL PBS. Wash plates five times with 10 mM PBS containing 1% Tween. LPS-Coated microtiter plates can be stored at room temperature for several months. 2. 1% containing 2% FBS (v/v). Use triplicate wells. Incubate at 37°C overnight.
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