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 language | English |
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Pages (from-to) | 19-26 |
Number of pages | 8 |
Journal | Journal of Immunological Methods |
Volume | 270 |
Issue number | 1 |
DOIs | |
State | Published - 1 Dec 2002 |
Keywords
- Adsorbance
- Detergent
- ELISA
- Insoluble
- SDS