EDTA和核黄素光聚合用于聚丙烯酰胺凝胶电泳分析

IF 1.9 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY The Protein Journal Pub Date : 2022-07-27 DOI:10.1007/s10930-022-10068-3
Volodymyr Shlyakhovenko, Olena Samoylenko
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引用次数: 1

摘要

提出了一种光敏聚合聚丙烯酰胺凝胶的新方法。以乙二胺四乙酸二钠盐(EDTA)和光引发剂核黄素(RF)为催化剂,催化丙烯酰胺/N,N′-亚甲基双丙烯酰胺(AM/Bis)的光聚合。制备的交联AM/Bis + EDTA/RF凝胶在高浓度AM (20 wt%)的电泳SDS-PAGE系统中进行检测。这些系统的效率电泳分离的人血淋巴细胞组蛋白被证明。原则上,这种在分离区具有小孔隙的凝胶可以为蛋白质的分解提供优势。该方法的优点还包括技术简单,可以及时制备凝胶(10-15 min)。然而,在微孔凝胶系统中,电印迹技术可能会出现一些限制,这对疏水蛋白尤其重要。
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Photopolymerization with EDTA and Riboflavin for Proteins Analysis in Polyacrylamide Gel Electrophoresis

A new method for photosensitized polymerization of polyacrylamide gels was proposed. Photopolymerization of acrylamide/N,N′-methylenebisacrylamide (AM/Bis) was assisted with combination of catalyst ethylenediaminetetraacetic acid disodium salt dihydrate (EDTA) and photoinitiator riboflavin (RF). The prepared cross-linked AM/Bis + EDTA/RF gels were tested in electrophoretic SDS–PAGE system at high concentration of AM (20 wt%). The efficiency of these systems for electrophoretic separation of histones of human blood lymphocytes was demonstrated. In principle, such gels with small pores in the separation zone can offer advantages for resolution of proteins. The advantages of proposed method also include simple technique and possibility of gel preparation in a timely manner (for 10–15 min). However, in microporous gel systems some limitations in electroblotting technique could occur, which is particularly crucial for hydrophobic proteins.

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来源期刊
The Protein Journal
The Protein Journal 生物-生化与分子生物学
CiteScore
5.20
自引率
0.00%
发文量
57
审稿时长
12 months
期刊介绍: The Protein Journal (formerly the Journal of Protein Chemistry) publishes original research work on all aspects of proteins and peptides. These include studies concerned with covalent or three-dimensional structure determination (X-ray, NMR, cryoEM, EPR/ESR, optical methods, etc.), computational aspects of protein structure and function, protein folding and misfolding, assembly, genetics, evolution, proteomics, molecular biology, protein engineering, protein nanotechnology, protein purification and analysis and peptide synthesis, as well as the elucidation and interpretation of the molecular bases of biological activities of proteins and peptides. We accept original research papers, reviews, mini-reviews, hypotheses, opinion papers, and letters to the editor.
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