Sodium dodecyl sulphate-treated proteins as ligands in ELISA

Valeria Lechtzier, Marina Hutoran, Talia Levy, Moshe Kotler, Talma Brenner, Michael Steinitz*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

20 Scopus citations

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.

Original languageEnglish
Pages (from-to)19-26
Number of pages8
JournalJournal of Immunological Methods
Volume270
Issue number1
DOIs
StatePublished - 1 Dec 2002

Keywords

  • Adsorbance
  • Detergent
  • ELISA
  • Insoluble
  • SDS

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