Institute of Nano- and Biotechnologies, Aachen University of Applied Sciences, 52428, Jülich, Germany.
Institute of Nano- and Biotechnologies, Aachen University of Applied Sciences, 52428, Jülich, Germany.
Anal Biochem. 2022 Oct 1;654:114819. doi: 10.1016/j.ab.2022.114819. Epub 2022 Jul 15.
An improved and convenient ninhydrin assay for aminoacylase activity measurements was developed using the commercial EZ Nin™ reagent. Alternative reagents from literature were also evaluated and compared. The addition of DMSO to the reagent enhanced the solubility of Ruhemann's purple (RP). Furthermore, we found that the use of a basic, aqueous buffer enhances stability of RP. An acidic protocol for the quantification of lysine was developed by addition of glacial acetic acid. The assay allows for parallel processing in a 96-well format with measurements microtiter plates.
开发了一种改良的、方便的用于测定氨基酰化酶活性的茚三酮法，使用的是商业 EZ Nin™试剂。还评估和比较了文献中的其他替代试剂。向试剂中添加 DMSO 可提高 Ruhemann 的紫（RP）的溶解度。此外，我们发现使用碱性水性缓冲液可增强 RP 的稳定性。通过添加冰醋酸开发了一种用于赖氨酸定量的酸性方案。该测定法允许在 96 孔格式中进行平行处理，并使用微量滴定板进行测量。
