Ni-NTA Fast Start Kit (6)

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Publication protocol

Protein isolation: Cultures were normalized by absorbance (600 nm) and fractionated using the ice-cold osmotic shock procedure [18], [46]. Western blotting of these fractions was performed [18]. Samples were read on a fluorescent microplate reader. For in vitro folding, native protein was purified on Ni-NTA columns according to the manufacturer's specifications (Ni-NTA Fast Start Kit, QIAGEN). Samples were concentrated and recovered in PBS buffer according to manufacturer's specifications (VivaSpin 6, Viva Science). SDS-PAGE was performed to verify binding efficiency. A Bio-Rad Protein Assay was used to quantify purified protein (Bio-Rad).

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