EC Number   |
Reference |
|---|
   7.5.2.6 | Ni-NTA column chromatography and DEAE column chromatography |
734183 |
| 7.5.2.6 | Ni2+ affinity resin column chromatography |
734221 |
| 7.5.2.6 | Ni2+-nitrilotriacetic acid resin column chromatography |
734250 |
| 7.5.2.6 | recombinant FLAG-tagged enzyme from Escherichia coli strain Rosetta 2 (DE3) by affinity chromatography, the Flag tag is removed using TEV protease, followed by gel filtration and ultrafiltration |
751693 |
| 7.5.2.6 | recombinant His10-tagged wild-type and mutant enzymes from Escherichia coli strain BL21-Codon plus(DE3)-RIL by solubilization from membranes with 2% n-dodecyl-beta-D-maltopyranoside and 0.4% cholate, cobalt affinity chromatography, and gel filtration in presence of detergents |
751037 |
| 7.5.2.6 | recombinant His6-tagged wild-type and mutant enzyme by nickel affinity chromatography |
696114 |
| 7.5.2.6 | recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli by ultraccentrifugation, detergent solubilization, again ultraccentrifugation, and nickel affinity chromatography, to over 95% purity |
751011 |
| 7.5.2.6 | recombinant His6-tagged wild-type and mutant enzymes from Lactobacillus lactis by soluibilization from membranes with detergent n-dodecyl beta-D-maltoside, and nickel affinity chromatography, formation of inside-out membrane vesicles |
696222 |
| 7.5.2.6 | recombinant N-terminal His6-tagged enzyme from Escherichia coli strain BL21 (DE3) by ultracentrifugation delivering inside-out membrane vesicles, solubilization by detergent, again ultracentrifugation, metal affinity chromatography, and desalting gel filtration. The suitable detergent for solubilization of MsbA is examined among a number of detergents including octylglucoside at 2% w/v, lauryldimethylamine oxide at 2% w/v, dodecyl beta-D-maltoside at 1% w/v and sarcosyl at 1% w/v. Samples are then incubated with gentle stirring for 1 h. No difference in the amount of solubilized proteins is observed when the membranes are incubated with detergents for longer than 1 h. Sarcosyl is most effective for solubilizing MsbA from the membranes resulting in the highest yield of solubilized protein compared with the other three detergents |
751020 |
| 7.5.2.6 | recombinant N-terminally His-tagged wild-type and mutant MsbAs from Escherichia coli, preparation of inside-out membrane vesicles, reconstitution of purified MsbA proteins in proteoliposomes, solubilization by 1% w/v n-dodecyl-beta-D-maltoside, and further purification by nickel affinity chromatography. The orientation of MsbA proteins in right-side-out membrane vesicles, inside-out membrane vesicles or proteoliposomes is assessed by determining the accessibility of the N-terminal His-tag to digestion by protease K in the external buffer |
751656 |
