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Photopolymerization with EDTA and Riboflavin for Proteins Analysis in Polyacrylamide Gel Electrophoresis

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Abstract

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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Fig. 1
Fig. 2

Data Availability

The data that support the findings of this study are available from the author, Olena Samoylenko (a-samoilenko@ukr.net) upon reasonable request.

Abbreviations

AM:

Acrylamide

APS:

Ammonium persulfate

Bis:

N,N′-methylenebisacrylamide

EDTA:

Ethylenediaminetetraacetic acid disodium salt dihydrate

LED:

Light-emitting diode

MB:

Methylene blue

PAGE:

Polyacrylamide gel electrophoresis

PMSF:

Phenylmethylsulfonylfluoride

RF:

Riboflavin

ROS:

Reactive oxygen species

SDS:

Sodium dodecyl sulfate

TEMED:

N,N,N′,N′-tetramethylethylenediamine

Tris:

Tris(hydroxymethyl)aminomethane

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Acknowledgements

The work was supported by the complex interdisciplinary research program of NAS of Ukraine, Projects # 0117U004977.

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  1. Laboratory of Metastatic Microenvironment Problems, RE Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology of the NASU, 45, Vasylkivska str, Kiev, 03022, Ukraine

    Volodymyr Shlyakhovenko & Olena Samoylenko

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  1. Volodymyr Shlyakhovenko
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Correspondence to Olena Samoylenko.

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Shlyakhovenko, V., Samoylenko, O. Photopolymerization with EDTA and Riboflavin for Proteins Analysis in Polyacrylamide Gel Electrophoresis. Protein J 41, 438–443 (2022). https://doi.org/10.1007/s10930-022-10068-3

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