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藻胆蛋白荧光标记物的光稳定性研究

Photostability studies of phycobiliprotein fluorescent labels.

作者信息

White J C, Stryer L

出版信息

Anal Biochem. 1987 Mar;161(2):442-52. doi: 10.1016/0003-2697(87)90473-8.

Abstract

Photostability studies of four fluorescent phycobiliproteins were conducted to identify stable chromophores for biological labeling applications. Phycobiliprotein photodestruction was linear with the applied laser power and depended on the total number of photons absorbed per molecule. Photodestruction quantum yields phi of 1.1 X 10(-5) for R-phycoerythrin, 6.6 X 10(-6) for B-phycoerythrin, 4.5 X 10(-6) for allophycocyanin, and 2.5 X 10(-6) for C-phycocyanin were measured. C-Phycocyanin is a factor of 10.8 more photostable than fluorescein. The photostability of R-phycoerythrin was improved by a factor of 1.7 by adding n-propyl gallate. The addition of superoxide dismutase, catalase, sodium dithionite, ascorbate, dithiothreitol, EDTA, or deoxygenation with argon bubbling had no effect on the photostability of R-phycoerythrin.

摘要

对四种荧光藻胆蛋白进行了光稳定性研究,以确定适用于生物标记应用的稳定发色团。藻胆蛋白的光破坏与所施加的激光功率呈线性关系,并取决于每个分子吸收的光子总数。测得R-藻红蛋白的光破坏量子产率φ为1.1×10^(-5),B-藻红蛋白为6.6×10^(-6),别藻蓝蛋白为4.5×10^(-6),C-藻蓝蛋白为2.5×10^(-6)。C-藻蓝蛋白的光稳定性比荧光素高10.8倍。通过添加没食子酸正丙酯,R-藻红蛋白的光稳定性提高了1.7倍。添加超氧化物歧化酶、过氧化氢酶、连二亚硫酸钠、抗坏血酸、二硫苏糖醇、乙二胺四乙酸或用氩气鼓泡进行脱氧处理对R-藻红蛋白的光稳定性没有影响。

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