How should the standard curve be prepared?
Despite the boiling of the samples there is some interference by IgG in the samples. That is why we recommend that the user adds antibodies to the standard curve in a similar concentration to that in the samples. The interaction between protein A and monoclonal antibodies varies depending on the class and subclass of the antibody. For many researchers it is probably sufficient to add any kind of monoclonal antibody to the samples, we recommend that the customer use the same antibody in the standard as in the samples. If the antibody used for the standard curve is purified on a protein A column it may contain some protein A thus adding protein A to the standard. It is therefore better to use an antibody in the standard curve that has been purified with another method than protein A. The antibody in the standard curve does not have to be purified, but the antibody concentration should be similar to the samples.
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- How should the standard curve be prepared?
