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Glutathione sepharose 4b resin

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Glutathione-Sepharose 4B resin is a chromatography matrix designed for the purification of glutathione-binding proteins. It consists of glutathione covalently coupled to Sepharose 4B beads. The resin can be used for the affinity purification of proteins that interact with glutathione, such as glutathione S-transferases.

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Lab products found in correlation

Anti phosphoserine antibody, by Merck Group (1 mentions) Ni nta agarose, by Qiagen (1 mentions)

2 protocols using glutathione sepharose 4b resin

1

Purification and Phosphorylation Analysis of Recombinant Proteins

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Ni-NTA agarose (Qiagen) and Glutathione-Sepharose 4B resin (Thermo Scientific) were used to purify the recombinant proteins for in vitro binding assays. NST1-GST and NST1(S316A)-GST fusion proteins were incubated with or without SnRK2s-6xHis fusion proteins in kinase buffer (50 mM Hepes [pH 7.4], 10 mM MgCl2, 1 mM dithiothreitol, 30 μM ATP) at 30 °C for 2 h. The reaction was stopped and divided into two equal portions. One group of proteins was separated by SDS/PAGE (Thermo-Fisher NuPAGE 4 to 12%, Bis-Tris, 1.5 mm, Mini Protein Gel, 10 well) and stained with Coomassie brilliant blue as the loading control. The other group was also separated by PAGE, and the gel was used for Western blotting with antiphosphoserine antibody (Sigma P5747) to detect phosphorylated proteins.
Liu C., Yu H., Rao X., Li L, & Dixon R.A. (2021). Abscisic acid regulates secondary cell-wall formation and lignin deposition in Arabidopsis thaliana through phosphorylation of NST1. Proceedings of the National Academy of Sciences of the United States of America, 118(5), e2010911118.
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2

Affinity Purification of Protein Complexes

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Cos-7 cells transfected with plasmids of interest were lysed with a lysis buffer (25 mM Tris [pH 7.4], 150 mM NaCl, 1% Triton X-100) and centrifuged at 16,000 × g for 10 min. The supernatant was incubated with glutathione Sepharose 4B resin (Thermo Fisher Scientific) for 30 min at 4°C with gentle agitation. After washing, the precipitated samples were subjected to immunoblot analysis.
Nishijima E., Honda S., Kitamura Y., Namekata K., Kimura A., Guo X., Azuchi Y., Harada C., Murakami A., Matsuda A., Nakano T., Parada L.F, & Harada T. (2022). Vision protection and robust axon regeneration in glaucoma models by membrane-associated Trk receptors. Molecular Therapy, 31(3), 810-824.
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