EC Number |
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1.13.12.6 | - |
1.13.12.6 | biotinylated enzyme is incubated with 1 microM sodium metaperiodate in 0.1 M sodium acetate buffer (pH 5.2), reaction stopped by glycerol addition, loading onto a size exclusion column, oxidized biotinylated enzyme eluted with the same buffer, pooling of bioluminescent fractions, concentration by ultrafree-0.5 centrifugal filter device, addition of 10 mM HiLyte Fluor 647 hydrazine in 0.1 M sodium acetate buffer (pH 5.2), after incubation loading onto a size-exclusion column, elution with 0.1 M potassium phosphate buffer (pH 7.2) containing 0.15 M NaCl. Treatment of human Delta-like protein (Dlk-1) antibody with 2-mercaptoethylamine (cleaving hinge-region disulfide bonds between heavy chains of antibody molecules) and conjugation of half antibodies to maleimide-activated avidin and purification on a size-exclusion column results in a 200 kDa conjugate |
1.13.12.6 | Escherichia coli cell culture with expression vector is cooled on ice-water bath, expression induced, incubation for 18 h at 15°C, cell concentration by centrifugation, solution in 50 mM Tris-HCl (pH 7.6) and 10 mM EDTA, disruption by sonication, centrifugation, SDS-PAGE |
1.13.12.6 | loading of recombinant prostaglandin E2 luciferase onto size-exclusion column, elution with 0.1 M potassium phosphate buffer, pH 7.2, and 0.15 M NaCl, fractions with bioluminescent activities (Cypridina luciferin solution, 0.1 M Tris-HCl, pH 7.4, 0.3 M sodium ascorbate, 0.2 M Na2SO3) are combined and stored at -30°C |
1.13.12.6 | native enzyme by fractional precipitation with acetone and ammonium sulfate, and dialysis, followed by gel filtration |
1.13.12.6 | native enzyme from photogenic organs by fractional precipitation with acetone and ammonium sulfate, and dialysis, followed by gel filtration |
1.13.12.6 | Ni affinity column chromatography, SA beads column chromatography, monomeric avidin column chromatography, and TSK G300SW gel filtration |
1.13.12.6 | recombinant secreted enzyme from Nicotiana benthamiana BY-2 cell medium by ultrafiltration, buffer exchange gel filtration, anion exchange chromatography, and dialysis |