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The procedure below provides a general guidance for the preparation of commonly tested samples for use in ELISA assays. Please check with the literature for experiments similar to yours for your new assay development. In this article, the following samples are covered:
Since serum tends to contain cytokines which may produce significant background signals, we suggest the preparation of serum-free or low-serum medium samples. If it is necessary to test serum containing medium, we also suggest running an uncultured medium blank to obtain the baseline signals which can then be subtracted from the cultured media sample data.
Guidelines on lysis buffer
Note: Hemolysis should be avoided while collecting serum samples. Those samples that have undergone hemolysis may increase non-specifically staining in HRP-conjugated ELISA assay.