NUSNNI-NanoCore, National University of Singapore, Singapore 117580, Singapore.
Anal Biochem. 2011 Oct 1;417(1):156-8. doi: 10.1016/j.ab.2011.05.045. Epub 2011 Jun 7.
This study revealed a major interference from sulfo-N-hydroxysuccinimide (sulfo-NHS) in the bicinchoninic acid (BCA) protein assay. Sulfo-NHS, a common reagent used in bioconjugation and analytical biochemistry, exhibited absorbance signals and absorbance peaks at 562 nm, comparable to bovine serum albumin (BSA). However, the combined absorbance of sulfo-NHS and BSA was not strictly additive. The sulfo-NHS interference was suggested to be caused by the reduction of Cu(2+) in the BCA Kit's reagent B (4% cupric sulfate) in a manner similar to that of the protein.
本研究揭示了磺基-N-羟基琥珀酰亚胺（sulfo-NHS）在二辛可宁酸（BCA）蛋白测定法中存在严重干扰。磺基-NHS 是生物偶联和分析生物化学中常用的试剂，在 562nm 处表现出吸光度信号和吸收峰，与牛血清白蛋白（BSA）相当。然而，磺基-NHS 和 BSA 的联合吸光度并非严格相加。磺基-NHS 的干扰被认为是由 BCA 试剂盒试剂 B（4%硫酸铜）中的 Cu(2+) 还原引起的，其方式类似于蛋白质。
