A simple and sensitive enzyme-mediated assay of biotin.
Free biotin was quantitated by a competition by coating biotin-bovine serum albumin conjugate on a polystyrene microplate for binding to avidin-beta-galactosidase conjugate. The enzyme conjugate remaining on the plate surface as a result of the competition was detected by reaction with one of the following fluorogenic substrates, resorufin beta-D-galactoside and fluorescein di-beta-D-galactoside, in a fluorescence plate reader. Free biotin as little as 0.1 nmol can be routinely detected.
Huang
Z
Z
Molecular Probes, Inc., Eugene, OR 97402.
Haugland
R P
RP
Szalecka
D
D
Haugland
R P
RP
D001360:Avidin; D001667:Binding, Competitive; D001710:Biotin / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D007124:Immunoenzyme Techniques; D013997:Time Factors; D001616:beta-Galactosidase
D001360:Avidin; D001710:Biotin; D001616:beta-Galactosidase