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Position: Discussion forums > Bioscience biotechniques discussion forums > Immunology forum and cytometry forum
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High Background Signal
Posted by: pmarino (IP Hidden, New member, 1)
Date: June 28, 2005 06:35AM
Good morning,
I am trying to develop a sandwich ELISA to detect Pichia pastoris host cell proteins. I have been running an experiment to detemine the levels of background noise. I have three control rows, where for each row, a different component is omitted and replaced with diluent buffer during incubation. The one row that is giving me trouble is the rown where the coating antibody is omitted. After the coating step, a blocking step is incorporated and the assay is run as a typical sandwich ELISA would be. I am getting a huge signal from this control well where the coating antibody is omitted. All other control wells are fine with regards to background noise. I was hoping a blocking step would eliminate this problem but it hasn't worked. Any suggestions at all in eliminate this issue would be extremely helpful.
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