Simple Western Streamlines Serum Antibody Analysis
Application Notes Summary
Scientists measure serum antibody levels to confirm immune responses against a bacteria to diagnose infections, to test for antibody production after vaccination, and to detect the presence of autoantibodies in autoimmune diseases. Traditional Western blots are often used to detect these antibodies, but testing with Western blots means a lot of hands-on time. After the antigen of interest is separated by SDS-PAGE and transferred to a membrane, each lane has to be cut into individual strips so patient serum samples can be individually tested for the presence of specific antibodies. Then you have to process and analyze them manually. Simple Western assays happen in individual capillaries, and everything from sample separation to data analysis is completely automated. No more cutting individual strips, washing and incubating them, or lining them all up with a molecular weight marker before detection. Just pipette your sample into the wells of your assay plate, set up your run, and you're done! And all that manual data analysis is gone too - Compass for Simple Western does it all for you. Did we mention you only need 10 µL of diluted serum per data point? That means you'll get a lot more data points for every 1 µL of neat serum. In this application note, we used Simple Western to detect autoantibodies in lupus patient serum as a model system to generate proof-of-concept data for the assay. But you can use this method any time you need to detect and quantitate specific serum antibodies. In fact, check out how researchers are using this assay to detect Salmonella antibodies without having to cut blots into individual strips.
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