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- Karlsson, Anna, 1967, et al.
(författare)
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Detection of glycoprotein receptors on blotting membranes by binding of live bacteria and amplification by growth.
- 1995
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Ingår i: Analytical biochemistry. - : Elsevier BV. - 0003-2697. ; 224:1, s. 390-4
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Tidskriftsartikel (refereegranskat)abstract
- Conditions have been adapted for detecting bacteria bound to glycoprotein receptors on blotting membranes using a self-enhancing detection method based on bacterial growth. Neutrophil plasma membrane proteins, mediating adherence of mannose-binding type-1-fimbriated Escherichia coli and concanavalin A (Con A) to intact human neutrophils, were separated by SDS-PAGE and transferred onto a polyvinylidene difluoride (PVDF) membrane. The PVDF membrane was immersed in a suspension of mannose-binding type-1-fimbriated E. coli, and after repeated washings, bound bacteria were allowed to multiply into bacterial colonies by placing the membrane on a solid nutrient substratum. About one major and eight minor glycoproteins, some of which also were detected by Con A, selectively induced colony formation in a mannose-inhibitable fashion. Binding of [35S]methionine metabolically labeled E. coli to PVDF membranes produced a virtually identical binding pattern, demonstrating further the accuracy of this self-enhancing detection method which is rapid, simple, and sensitive and avoids radioisotopes.
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