Fluorescent Labeling of DR5-B-iRGD Protein

To obtain fluorescently labeled protein DR5-B-iRGD, first the DR5-B-iRGD amino acid sequence was genetically modified at the N-end by replacing the amino acid residue valine in the 114 position to cysteine by site-directed mutagenesis. The cysteine-modified proteins DR5-B and DR5-B-iRGD were obtained by the method earlier reported for DR5-B [45 (link)]. Further, the cysteine-modified proteins DR5-B and DR5-B-iRGD were labeled by maleimide chemistry coupling with the fluorescent dye sulfo-Cyanine 3 maleimide (Lumiprobe, Moscow, Russia) according to the manufacturer’s protocol.

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Yagolovich A.V., Isakova A.A., Artykov A.A., Vorontsova Y.V., Mazur D.V., Antipova N.V., Pavlyukov M.S., Shakhparonov M.I., Gileva A.M., Markvicheva E.A., Plotnikova E.A., Pankratov A.A., Kirpichnikov M.P., Gasparian M.E, & Dolgikh D.A. (2022). DR5-Selective TRAIL Variant DR5-B Functionalized with Tumor-Penetrating iRGD Peptide for Enhanced Antitumor Activity against Glioblastoma. International Journal of Molecular Sciences, 23(20), 12687.

Publication 2022

sulfo-Cyanine 3 maleimide

Amino acid Amino acid sequence Cysteine Fluorescent dye Maleimide Proteins b Site directed mutagenesis Valine

Corresponding Organization : Lomonosov Moscow State University

Other organizations : P.A. Hertzen Moscow Oncology Research Institute, Ministry of Health of the Russian Federation

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Variable analysis

independent variables

Genetic modification of DR5-B-iRGD amino acid sequence at the N-end to replace valine in the 114 position with cysteine by site-directed mutagenesis

dependent variables

Obtaining cysteine-modified proteins DR5-B and DR5-B-iRGD

control variables

The method previously reported for obtaining DR5-B [45]

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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