Streptavidin protein

SDS-PAGE was performed using 18% polyacrylamide gel under reducing condition. The separated proteins were transferred to PVDF membranes (Merck Millipore). After blocking the membrane with 5% skim milk, apoC-II, apoC-III, or apoA-I was primarily detected with goat anti-apoC-II, -apoC-III, or -apoA-I polyclonal antibody (Academy Bio-Medical Company), respectively, and horseradish peroxidase (HRP)-conjugated rabbit anti-goat IgG antibody (Medical & Biological Laboratories) was the secondary antibody. Finally, the band containing apoC-II or apoC-III was visualized using ECL Prime Western Blotting Detection Reagent (GE Healthcare), and the apoA-I band was visualized using 3,3′-diaminobenzidine tetrahydrochloride and hydrogen peroxide. In the case of biotinylated proteins, the detection was performed similarly except that 5% BSA was used as a blocking solution and detection was carried out using Streptavidin Protein, HRP (Thermo Fisher scientific), and ECL Prime Western Blotting Detection Reagent.

NovaSyn TGR resin was purchased from Novabiochem (Darmstadt, Germany). Dichloromethane (DCM) and trifluoro acetic acid (TFA) were obtained from Watanabe Chemical Industries (Hiroshima, Japan). N,N-Dimethylformamide (DMF) was obtained from Kanto Chemical (Tokyo, Japan). Chloroacetyl chloride and triisopropylsilane (TIS) were obtained from Tokyo Chemical Industry (Tokyo, Japan). Dimethyl sulfoxide (DMSO) was obtained from Wako Pure Chemical Industries (Osaka, Japan). Methyl-β-cyclodextrin (M-β-CD) was obtained from Sigma Aldrich (St. Louis, MO). NHS-rhodamine (TAMRA SE) and streptavidin protein were obtained from Thermo Fisher Scientific (Waltham, MA).

All lipid stocks that we used for the experiments were prepared in chloroform and used as received (Avanti Polar Lipids and Echelon Biosciences). Alexa Fluor 488 conjugated This journal is © The Royal Society of Chemistry 2022 streptavidin protein (ThermoFisher Scientific) and 50 wt% polyethyleneimine solution (PEI, MW 2000) were procured from Sigma Aldrich. In all our experiments, we used freshly prepared Tris-buffer (pH 7.4). The various lipids are tabulated in Table 1.