The FITC-anti-FITC system is a sensitive alternative to biotin-streptavidin in ELISA

Harmer, I. and Samuel, D. 1989. The FITC-anti-FITC system is a sensitive alternative to biotin-streptavidin in ELISA. Journal of Immunological Methods. 122 (1), pp. 115-121. doi:10.1016/0022-1759(89)90341-4

TitleThe FITC-anti-FITC system is a sensitive alternative to biotin-streptavidin in ELISA
AuthorsHarmer, I. and Samuel, D.
Abstract

In a model ELISA system alkaline phosphatase (AP) absorbed onto microtitre wells was employed as the target antigen. The antigen was then reacted with a monoclonal antibody to AP either unlabelled or labelled with (a) FITC and (b) biotin. The bound anti-AP was then detected with horseradish peroxidase (HRP) conjugates of polyvalent anti-mouse IgG, and the FITC and biotin anti-AP conjugates with HRP conjugates of a monoclonal anti-FITC or streptavidin respectively. The FITC-anti-FITC system proved to be of similar sensitivity to the biotin-streptavidin system detecting 140 amol compared to 350 amol of antigen. Both these methods of antigen detection were superior to the anti-mouse IgG reagent (2100 amol). In contrast to biotinylated antibodies, FITC-labelled antibodies are highly coloured and fluorescent. These features aid the preparation, purification and characterisation of conjugates. In addition, very low non-specific binding is encountered with enzyme conjugates of anti-FITC and this may confer an advantage over enzyme conjugates of avidin/streptavidin reagents.

JournalJournal of Immunological Methods
Journal citation122 (1), pp. 115-121
ISSN0022-1759
Year1989
PublisherElsevier
Digital Object Identifier (DOI)doi:10.1016/0022-1759(89)90341-4
Publication dates
Published15 Aug 1989

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