- Membrane strips containing electrophoretically separated antigen extracts are used as solid phase. The position of the proteins depends on their respective molecular masses.
- If the sample is positive, specific antibodies in the diluted serum sample attach to the antigens coupled to the membrane.
- In a second incubation step, the attached antibodies react with AP-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.
- Evaluating the band patterns on the incubated membrane strips involves differentiating non-specific from specific antibodies. The number and intensity of the specific bands is decisive for the result "positive/negative".
- EUROLINE-WB: detection of antibodies against Borrelia.
- A separate membrane strip is incubated for each serum sample.
- Total time for performing the Westernblot test is about 115 minutes.
- All incubation steps proceed at room temperature.
- The bands are assigned according to a lot-specific evaluation matrix provided. A separate lot is issued for each electrophoresis gel, helping to avoid errors in the assignment of the bands.
- Every test kit contains a membrane strip of the same lot incubated with a positive reference serum. Therefore, there is no need to incubate a positive control serum.
- The membrane strips are pre-numbered to prevent confusion. Laborious labelling is not necessary.
- Correct completion of the individual incubation steps for each membrane strip is indicated by staining of the control band at the bottom of the strip.
- Positive and negative reactions can be easily and reliably differentiated from each other. The intensity of the antigen bands correlates with the antibody titer.
- The Westernblot is the method of choice when the objective is to confirm or differentiate positive results obtained in a screening test (indirect immunofluorescence or microplate ELISA).
- EUROLINE-WB is a combination of westernblot and line blot techniques. Proteins from a whole antigen extract are electrophoretically separated according to molecular mass and transferred onto a nitrocellulose membrane. Highly purified native or recombinant antigens are then printed as lines onto the westernblot strips (EUROLINE membrane chip).
- The program EUROLineScan from EUROIMMUN has been developed to enable quantitative evaluation of Westernblot/EUROLINE-WB test strips, to facilitate management of data, and to provide detailed documentation of results. First, the incubated Westernblot/EUROLINE-WB 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.
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