Fig. 4

Purification of 6h-Aadh2p by Ni–NTA. a Coomassie-stained SDS-PAA gel and western Blot from crude extract (1) and eluate 1 (2) fractionated by electrophoresis on 12 % gel. The primary antibody was anti-poly Histidine from mice and the secondary antibody was anti-mouse IgG alkaline phosphatase. b Summary of 6h-Aadh2p purification. Intracellular soluble fraction of disrupted yeast cells from strain G1212/YIC102-6H-AADH2 was named ‘Crude extract’, proteins that did not bind to Ni–NTA were called ‘Flow-through’, proteins that were washed from the material was named ‘Wash’ and proteins that were eluted called ‘Eluate 1 and ‘Eluate 2′. Activity (A) was determined as described in the assay for determination of ADH activity