ScFv16 was expressed in High-Five™ insect cells (ThermoFisher Scientific, Cat#B85502) as a secreted protein purified by Ni-sepharose chromatography column49 (link). The HiLoad 16/600 Superdex 75 column (GE Healthcare) was used to separate the monomeric fractions of scFv16 with a running buffer containing 20 mM HEPES and 100 mM NaCl, pH 7.4. The purified scFv16 was flash-frozen in liquid nitrogen with 10% glycerol and stored at −80 °C until use.
Chen Y., Zhou Q., Wang J., Xu Y., Wang Y., Yan J., Wang Y., Zhu Q., Zhao F., Li C., Chen C.W., Cai X., Bathgate R.A., Shen C., Eric Xu H., Yang D., Liu H, & Wang M.W. (2023). Ligand recognition mechanism of the human relaxin family peptide receptor 4 (RXFP4). Nature Communications, 14, 492.
BufferCells Frozen GlycerolHepes Insect NaclNitrogen Protein Sepharose chromatography
Corresponding Organization :
Other organizations :
Fudan University, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Institute of Deep-Sea Science and Engineering, Florey Institute of Neuroscience and Mental Health, University of Melbourne
Running buffer containing 20 mM HEPES and 100 mM NaCl, pH 7.4
Freezing of purified scFv16 in liquid nitrogen with 10% glycerol and storage at -80 °C
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