超滤作为牛血清白蛋白-异硫氰酸荧光素生物偶联物的替代纯化方法。

microPublication biology Pub Date : 2025-07-23 eCollection Date: 2025-01-01 DOI:10.17912/micropub.biology.001731
Catherine A Jackson, Elise L McKean, John M Hawdon
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引用次数: 0

摘要

蛋白质与荧光探针的生物偶联是一种流行的蛋白质可视化方法,在科学领域有着广泛的应用。本研究评估了超滤作为异硫氰酸荧光素(FITC)与牛血清白蛋白(BSA)标记生物偶联反应后的一种替代纯化方法。通过(1)紫外可见光谱法,将样品与市售产品进行比较,以及(2)在体外生物测定中使用,确定了纯化的成功。离心式超滤装置的分子量低于共轭蛋白的分子量,可以成功地保留纯化产物,并从溶液中清除未反应的FITC。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Ultrafiltration as an alternative purification method for bovine serum albumin-fluorescein isothiocyanate bioconjugate.

Ultrafiltration as an alternative purification method for bovine serum albumin-fluorescein isothiocyanate bioconjugate.

Ultrafiltration as an alternative purification method for bovine serum albumin-fluorescein isothiocyanate bioconjugate.

The bioconjugation of proteins with fluorescent probes is a popular method for protein visualization and has widespread applications across scientific fields. This study assessed ultrafiltration as an alternative purification method following the bioconjugation reaction that tags fluorescein isothiocyanate (FITC) to bovine serum albumin (BSA). Success of the purification was determined by (1) UV-Vis spectroscopy, comparing our sample to a commercially available product, and (2) use in an in vitro bioassay. Centrifugal ultrafiltration devices with a molecular weight cut off below the molecular weight of the conjugated protein were successful in retaining the purified product and clearing unreacted FITC from solution.

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