Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 417394
Title A duplex approach for immunochemical staining and typing of protein in western blots
Author(s) Kuczius, T.; Brandstädter, L.; Karch, H.; Langeveld, J.P.M.
Source Analytical Biochemistry 409 (2011)2. - ISSN 0003-2697 - p. 260 - 266.
DOI http://dx.doi.org/10.1016/j.ab.2010.10.018
Department(s) CVI Infection Biology
Publication type Refereed Article in a scientific journal
Publication year 2011
Keyword(s) cellular prion proteins - bovine spongiform encephalopathy - electrophoretic transfer - nitrocellulose - scrapie - peroxidase - brain - prpsc - visualization - sensitivity
Abstract The qualitative and semiquantitative Western blotting technique enables the detection of separate proteins and the determination of subtypes and fragments by specific immunological reactions. Protein typing on immunoblots is restricted to antibody-specific determination, with the result of a specific banding pattern. For protein characterization, several antibodies that recognize different epitopes within the protein sequence are used. However, repeated or parallel gel runs are needed. Here we describe a sequential determination of prion proteins in healthy and pathological states that both consist of di-, mono-, and nonglycosylated isoforms using a single blot with two antibodies from two species that recognize one antigen with two epitopes. The band signals are visualized by using different chemiluminescent substrate reactions. This application can be used in the fields of diagnostics and public health to detect full-length and fragmented proteins and can also be used for characterization of overlaying proteins.
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