Materials
Name | Company | Catalog Number | Comments |
Transmission electron microscope: JEOL2100F | JEOL | ||
CCD camera | Tiez Video and Imaging Processing System GmbH, Germany | ||
Glow discharger | Baltec | ||
TEM grid: 400 mesh | TAAB | GM016/C | |
Size exclusion chromatography: Agilent SEC-5 | Agilent Technologies | 5190-2526 | |
Superdex 200 HR 10/300 | GE Healthcare Life Sciences | 17-5172-01 | |
Plasmid: MSP1E3D1 | Addgene | 20066 | |
Bacteria: BL21DE3 | NEB | C2527H | |
Bacteria: BL21 (DE3) T1R pRARE2 | Protein Science Facility, KI, Solna | ||
Purification Matrix: ATP agarose | Sigma Aldrich | A2767 | |
Purification Matrix: HisTrap HP-5 mL | GE Healthcare Life Sciences | 17-5247-01 | |
Lipid: POPC | Avanti polar lipids | 850457C | 25 mg/mL in chloroform |
Hydrophobic beads: Bio-Beads, SM-2 Resin | Bio-Rad | 1523920 | |
13 mm syringe filter: 0.2 μm | Pall life sciences | PN 4554T | |
Stain: Sodium phosphotungstate tribasic hydrate | Sigma Aldrich | 31648 | |
2-mercaptoethanol | Sigma Aldrich | M3148-250ML | |
Sodium Dodecyl Sulfate (SDS) | Bio-Rad | 161-0301 | |
Protease inhibitor cocktail | Sigma Aldrich | 4693132001 | |
TCEP | Sigma Aldrich | 646547 | |
Detergent: Sodium cholate hydrate | Sigma Aldrich | C6445-10G | |
Sodium Cholate | 500 mM Sodium cholate. Resuspend in miliQ water and store at -20 °C. | ||
Lipid Stock | 50 mM POPC, 100 mM sodium cholate, 20 mM Tris-HCl pH 7.5, 100 mM NaCl. Store -80 °C for a month, after purging the solution with nitrogen. |
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MSP standard buffer | 20 mM Tris-HCl, pH 7.5, 100 mM NaCl, 0.5 mM EDTA. Store at 4 °C. |
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Non-Denaturaing Electrophoresis Anode Buffer | Thermo Fisher Scientific | BN2001 | 50 mM Bis-Tris, 50 mM Tricine, pH 6.8 |
Non-Denaturaing Electrophoresis Cathode Buffer | Thermo Fisher Scientific | BN2002 | 50 mM Bis-Tris, 50 mM Tricine, pH 6.8, 0.002% Coomassie G-250 |
Non-Denaturaing Electrophoresis 4x Sample loading Buffer | Thermo Fisher Scientific | BN2003 | 50 mM Bis-Tris, pH 7.2, 6 N HCl, 50 mM NaCl, 10% (w/v) glycerol, 0.001% Ponceau S |
Denaturaing Electrophoresis Running Buffer | In-house recipe: 25 mM Tris-HCl, pH 6.8, 200 mM Glycine, 0.1% (w/v) SDS | ||
Denaturaing Electrophoresis 5x Sample loading Buffer | In-house recipe: 0.05% (w/v) Bromophenolblue, 0.2 M Tris-HCl, pH 6.8, 20% (v/v) glycerol, 10% (w/v) SDS, 10 mM 2-mercaptoethanol | ||
Terrific broth | Tryptone - 12.0 g, Yeast Extract - 24.0 g, 100 mL 0.17 M KH2PO4 and 0.72 M K2HPO4, Glycerol - 4 mL. Tryptone, yeast extract and glycerol were prepared to 900 mL and autoclaved seperately. KH2PO4 and K2HPO4 were prepared and autoclaved separately. Both were mixed before using the medium. |
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