Huang Z, Haugland R P, Szalecka D, Haugland R P
Molecular Probes, Inc., Eugene, OR 97402.
Biotechniques. 1992 Oct;13(4):543-6.
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.
通过将生物素 - 牛血清白蛋白偶联物包被在聚苯乙烯微孔板上以与抗生物素蛋白 - β - 半乳糖苷酶偶联物结合的竞争反应来定量游离生物素。竞争反应后残留在板表面的酶偶联物通过与以下荧光底物之一，试卤灵β - D - 半乳糖苷和荧光素二β - D - 半乳糖苷反应，在荧光酶标仪中进行检测。常规情况下可检测低至0.1 nmol的游离生物素。
