EC Number   |
|---|
   2.4.99.18 | - |
| 2.4.99.18 | affinity purification of a tagged Stt3 protein (component of the oligosaccharyltransferase complex) |
| 2.4.99.18 | C-terminal domain of Stt3p, wild-type and mutant D518E |
| 2.4.99.18 | cells centrifuged, resuspended, and washed in 20 mM phosphate buffer, pH 7.2, with 0.3 M Na Cl, cells disrupted and centrifuged, membranes separated by ultracentrifugation of supernatant, resuspended in buffer with 2% Elugent and 25 mM imidazole buffer, proteins solubilized by tumbling, centrifuged, supernatant removed, Elugenat reduced to 1% or 0.5% n-dodecyl-beta-D-maltopyranoside, loaded onto Ni-nitrilotriacetic acid agarose column, elution with buffer containing 250 mM imidazole |
| 2.4.99.18 | dissolution in denaturing buffer (6 M guanidine hydrochloride, 500 mM Na Cl, 25 mM imidazole, 20 mM phosphate buffer, pH 7.4), cenrtrifugation, supernatant loaded onto nickel-NTA column for affinity chromatography with 20 mM phosphate buffer, pH 6.5, SDS-PAGE |
| 2.4.99.18 | Ni-NTA resin column chromatography |
| 2.4.99.18 | partial |
| 2.4.99.18 | purification of an affinity-tagged version of the enzyme complex from a membrane protein fraction |
| 2.4.99.18 | recombinant N-terminally His6-tagged wild-type and selenomethionine-labeled AglB from Escherichia coli strain BL21 by nickel affinity chromatography and gel filtration, followed by anion exchange chromatography |
| 2.4.99.18 | sonication, centrifugation, supernatant absorbed to glutathione-Sepharose 4B resin, elution, concentration, reductive methylation, gel filtration with Superdex75 column, followed by anion exchange chromatography with a Resource Q column |
