The monoclonal antibody 11E10 recognizes the 2A subunit of Shiga-like toxin 2. Shiga-like toxins (SLTs), are also called Verotoxins. Enterohemorrhagic Escherichia coli (EHEC) strains which are primarily of serotypes 0157:H7, 026:H11, and O111:H8 have been incriminated as etiologic agents of hemorrhagic colitis and Hemolytic-uremic syndrome, a generalized disease characterized by acute renal failure, thrombocytopenia, and microangiopathic hemolytic anemia. There are several distinct E.coli SLTs. SLT-I and SLT-II are produced by EHEC. SLT-I and Shiga toxin share;99% deduced amino acid sequence homology, whereas SLT-I and SLT-II share about 60% deduced amino acid sequence homology. SLT-I and SLT-II are antigenically distinct. Antibodies to SLT-II can also neutralize a variant of SLT-II (designated SLT-Iiv) produced by strains of E.coli that cause edema. SLT-IIv is cytotoxic for Vero but not HeLa cells, distinguishing it from SLT-II. The protein structure of the toxin consists of two domains: the A polypeptide that inhibits protein synthesis by targeting ribosomes, and the B polypeptide pentamer that binds to the eukaryotic cell receptor globotriaosylceramide (Gb3) leading to receptor-mediated endocytosis.
Functional studies, Immuno assays, Western blot
For Western blotting dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:10. For inhibition of biological activity in vitro dilutions have to be made according to the amounts of toxin to be inactivated.