A duplex approach for immunochemical staining and typing of protein in western blots

T. Kuczius, L. Brandstädter, H. Karch, J.P.M. Langeveld

Research output: Contribution to journalArticleAcademicpeer-review

2 Citations (Scopus)

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.
Original languageEnglish
Pages (from-to)260-266
JournalAnalytical Biochemistry
Volume409
Issue number2
DOIs
Publication statusPublished - 2011

Keywords

  • cellular prion proteins
  • bovine spongiform encephalopathy
  • electrophoretic transfer
  • nitrocellulose
  • scrapie
  • peroxidase
  • brain
  • prpsc
  • visualization
  • sensitivity

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