PMID: 2503563Aug 15, 1989Paper

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

Journal of Immunological Methods
I J Harmer, D Samuel

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.

References

Aug 1, 1979·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·J L GuesdonS Avrameas
Jul 1, 1989·Journal of Bioluminescence and Chemiluminescence·D Ireland, D Samuel
May 15, 1988·Analytical Biochemistry·M Wilchek, E A Bayer
Mar 16, 1988·Journal of Immunological Methods·D SamuelR A Abuknesha
Jan 21, 1985·Journal of Immunological Methods·P ViljaP Tuohimaa
Nov 1, 1969·Archives of Biochemistry and Biophysics·M Steinbuch, R Audran

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Citations

Oct 4, 1990·Journal of Immunological Methods·D Samuel, R A Abuknesha
Feb 28, 1994·Journal of Immunological Methods·P BaloghP Németh
Nov 1, 1989·Journal of Virological Methods·K E BrownD Samuel
Jan 1, 1989·Current Opinion in Immunology·B Nilsson
Aug 1, 1990·Journal of Clinical Pathology·R J BirtlesA G Taylor
Mar 3, 1999·Proceedings of the National Academy of Sciences of the United States of America·G BesteA Skerra
Aug 10, 2007·Biochimie·Jörg A SchenkBurkhard Micheel
Jul 10, 2018·Journal of Immunoassay & Immunochemistry·Sasiwan KerdpooThanusak Tatu
Jul 9, 1999·Journal of Molecular Recognition : JMR·V BöttgerB Micheel
Jan 25, 2021·Advanced Drug Delivery Reviews·Pere MongeAlexander N Zelikin

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