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Technology

Blot Techniques

The EUROLINE/Western Blot: A New Technique for Extensive Antibody Profiles

Principle of the Test

  • Membrane strips coated with thin parallel lines of several purified, biochemically characterized antigens are used as solid phase. The membranes are fixed as onto synthetic foil.
  • If the sample is positive, specific antibodies in the diluted serum sample attach to the antigens coupled to the solid phase.
  • In a second incubation step, the attached antibodies react with alkaline-phosphatase-labelled anti-human antibodies.
  • In a third step, the bound antibodies are stained with a chromogen/substrate solution which is capable of promoting a color reaction. An intense dark band at the line of the corresponding antigen appears if the serum sample contains specific antibodies.
  • Depending on the spectrum of antigens used, it is possible to analyze several antibodies next to each other and simultaneously under identical conditions.
    • Easy Handling, Reliable and Simple Evaluation

      • A separate membrane strip is incubated for each serum sample.
      • Total time for performing the analysis is about 105 minutes.
      • The incubation can be automated using the EUROBlotMaster.
      • All incubation steps proceed at room temperature.
      • The antigen bands are located at exactly defined positions, which means that the evaluation of the test is much simpler than for Westernblots.
      • Correct completion of the individual incubation steps is indicated by staining of the control band on each EUROLINE test strip.
      • Positive and negative results can be easily and reliable differentiated from each other. The intensity of the antigen bands correlates with the antibody titer.
      • The antigens used are highly pure, mostly isolated by affinity chomatography. The membrane strips do not contain any superfluous proteins which might cause unspecific positive results.
      • The incubated EUROLINE test strips can be stored for long periods. Results can be easily documented.
      • The program EUROLineScan from EUROIMMUN has been developed to enable quantitative evaluation of EUROLINE test strips, to facilitate management of data, and to provide detailed documentation of results. First, the incubated EUROLINE test strips are scanned using a flatbed scanner. EUROLineScan recognizes the position of the strips, even if they have been placed inexactly, identifies the bands, and measures their intensity. The results are then saved together with the image data. A separate results sheet can be produced for each patient.