n-Octylpolyoxyethylene

This plan only provides a guide, please modify it to meet your specific needs.
1. Solution preparation
(1) Working solution: Dissolve n-Octylpolyoxyethylene in DMSO and dilute the stock solution with experimental buffer to the required working concentration.
Note: Please adjust the optimal working concentration according to the actual situation or refer to the literature to set the gradient concentration yourself. The working solution must be prepared and used immediately.

2. Protocol for refolding, purifying and reconstructing proteins using n-Octylpolyoxyethylene^[1] (Source literature, for reference only)
(1) Transform the expression plasmid containing the selected cDNA sequence into the Escherichia coli strain. After induction, high-pressure homogenization and centrifugation, the protein is separated into inclusion bodies.
(2) Dissolve 1mg of inclusion bodies in TE/G buffer containing 2% N-lauroylsarcosine, 1.3% Triton X-114, 0.3% n-Octylpolyoxyethylene, 1mM DTT and GTP (pH 7.5). Gradually mix in 50 mg of lipid mixture (DOPC:DOPE:CL, 45:45:10mol%).
(3) Concentrate the mixture and dialyze into the experimental buffer (50mM Na₂SO₄, 10mM MES, 10mM Tris and 0.6mM EGTA (pH 7.34)).
(4) Remove unfolded and aggregated proteins from the dialysate by centrifugation and a column containing hydroxyapatite. Use Bio-Beads SM-2 to remove nonionic detergents.
(5) Measure the protein concentration in the proteoliposomes using a protein assay kit. Verify protein purity by SDS-PAGE and silver staining.
Note: For your safety and health, please wear a lab coat and disposable gloves when operating.
References:
[1]Žuna K, Jovanović O, Khailova L S, et al. Mitochondrial uncoupling proteins (UCP1-UCP3) and adenine nucleotide translocase (ANT1) enhance the protonophoric action of 2, 4-dinitrophenol in mitochondria and planar bilayer membranes[J]. Biomolecules, 2021, 11(8): 1178.