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Size-exclusion chromatography multi-angle light scattering

EM Emelia Melvin
ZK Zuzana Kalaninová
ES Elia Shlush
PM Petr Man
MG Moshe Giladi
YH Yoni Haitin
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Experiments were performed using an analytical SEC column (Superdex 200 Increase 10/300 GL; Cytiva) preequilibrated with gel filtration buffer (20 mM Tris, pH 7.5, 150 mM NaCl, 0.3 mM DDM). Samples of purified mTTYH1-GFP-8xHis (50 μL, 1.18 mg/mL) were loaded onto the SEC column connected in line to a downstream 8-angle light scattering detector, followed by a differential refractive index detector (Wyatt Technology). Protein conjugate analysis was performed using a dedicated program within the Astra software package (Wyatt Technology) to determine the molecular mass of the protein and detergent components of the complex.

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