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  • Title: Sodium dodecyl sulphate-treated proteins as ligands in ELISA.
    Author: Lechtzier V, Hutoran M, Levy T, Kotler M, Brenner T, Steinitz M.
    Journal: J Immunol Methods; 2002 Dec 01; 270(1):19-26. PubMed ID: 12379335.
    Abstract:
    Insoluble proteins generally do not adsorb to microtitre wells and, therefore, cannot be used as antigens in enzyme-linked immunosorbent assay (ELISA). However, denaturation and solubilization with 2% sodium dodecyl sulphate (SDS) renders these proteins suitable ligands for ELISA. In quantitative ELISA using polyclonal antibodies as primary antibody, comparable results were obtained with native and SDS-denatured protein ligands. The binding of the antibodies to the SDS-treated ligands was completely inhibited by premixing the primary antibody with the corresponding native antigen. Nonspecific binding of primary and secondary antibodies to SDS-treated ligands was not observed. SDS-treated proteins are able to attach to ELISA microwells, retain their antigenic epitopes and do not engender an elevated background. The concentration of SDS-treated proteins required for coating is the same as that of the native proteins.
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