N nonyl β d glucoside

The detergent n-dodecyl-β-D-maltoside (DDM) was purchased from Inalco Pharmaceuticals (San Luis Obispo, CA), whereas all other detergents used were from Anatrace (Maumee, OH): n-undecyl-β-D-maltopyranoside (UDM), n-decyl-β-D-maltopyranoside (DM), n-octyl-β-D-glucoside (OG), n-nonyl-β-D-glucoside (NG), 3[(3-cholamidopropyl) dimethylammonio] propanesulfonic acid (CHAPS), n-dodecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate (AZ), 2,2-dioctylpropane-1,3-bis-β-D-maltopyranoside (DMNG), and n-dodecylphosphocholine (FS-12). For solubilization analysis, 10 μL of detergent mixture (2X of desired concentration (w/v)) dissolved in 500 mM NaCl, 50 mM Tris/HCl, pH 8, with 10% glycerol, was combined with 10 μl of membrane vesicles (4 mg/ml) diluted in the same buffer. The mixture was incubated at 4°C for 2 h with gentle rotation, followed by centrifugation at 125,000 g for 45 min to separate solubilized from unsolubilized material. Samples were subjected to SDS-PAGE and Western blot analysis as described earlier. ImageJ software was used for quantitative analysis.

Membranes were diluted with buffer A and mixed with different detergents to a final protein concentration of 2 mg mL⁻¹ and a detergent concentration of 10 × critical micelle concentration (CMC) [5.3% n-Octyl-β-D-glucoside (OG), 2% n-nonyl-β-D-glucoside (NG), 0.47% n-dodecylphosphocholine (Fos-choline-12, FC-12), and 0.087% n-dodecyl-β-D-maltopyranoside (DDM) (Anatrace, Maumee, Ohio, USA)]. The non-solubilized and solubilized proteins were separated (150,000× g, 30 min, 4 °C) and checked through Coomassie and Western-Blot.