-transparent.png){kind=logo}
Reciprocal enzyme assays are described for the vitamin biotin and for the biotin-binding proteins avidin and streptavidin. The assays are based on the following steps: (a) biotinylated bovine serum albumin is adsorbed onto microtiter plates; (b) streptavidin (or avidin) is bound to the biotin-coated plates; (c) biotinylated enzyme (in this case alkaline phosphatase) is then interacted with the free biotin-binding sites on the immobilized protein. For biotin assay, competition between the free vitamin and the biotinylated enzyme is carried out between steps (b) and (c). The method takes advantage of the four biotin-binding sites which characterize both avidin and streptavidin. The method is extremely versatile and accurate over a concentration range exceeding three orders of magnitude. The lower limits of detection are approximately 2 pg/ml (0.2 pg/sample) for biotin and less than 100 ng/ml (10 ng/sample) for either avidin or streptavidin.
E A Bayer, and
H Ben-Hur, and
M Wilchek
August 1977,
Analytical biochemistry,
E A Bayer, and
H Ben-Hur, and
M Wilchek
January 1979,
Methods in enzymology,
E A Bayer, and
H Ben-Hur, and
M Wilchek
January 1985,
Journal of immunological methods,
E A Bayer, and
H Ben-Hur, and
M Wilchek
January 1990,
Methods in enzymology,
E A Bayer, and
H Ben-Hur, and
M Wilchek
January 1988,
Journal - Forensic Science Society,
E A Bayer, and
H Ben-Hur, and
M Wilchek
January 2016,
PloS one,
E A Bayer, and
H Ben-Hur, and
M Wilchek
March 1986,
Journal of immunological methods,
E A Bayer, and
H Ben-Hur, and
M Wilchek
July 1993,
Nuclear medicine and biology,
E A Bayer, and
H Ben-Hur, and
M Wilchek
January 1964,
Biochemical and biophysical research communications,
E A Bayer, and
H Ben-Hur, and
M Wilchek
June 1965,
Biochemical and biophysical research communications,
