Journal: Protein expression and purification

Article Title: High yield bacterial expression and purification of active recombinant PA28alphabeta complex.

doi: 10.1016/j.pep.2008.10.014

Figure Lengend Snippet: Fig. 3. (A) Purification scheme of recombinant PA28ab complex. Details of the purification procedure are described under Materials and methods. (B) Final step of purification: gel filtration on Superose 12. Details of the chromatography are described under Materials and methods. Fractions 5–24 were incubated with purified 20S proteasomes for 20 min. at 30 C, then assayed on peptidase activity, using Suc-LLVY-AMC as a substrate (red dotted line). Blue line: optical density at 280 nm. The black arrowheads indicate the position of MW markers: from left to right, Ferritin (440 kDa), Aldolase (158 kDa), Ovalbumin (43 kDa), Chymotrypsinogen (25 kDa). (C) SDS–PAGE analysis of the peak fractions. Coomassie blue stained 12% high resolution SDS–PAGE gel of 10 lL samples of fractions 12, 13 and 14. (For interpretation of color mentioned in this figure the reader is referred to the web version of the article.)

Article Snippet: All chromatographic steps were performed using the Biological Duo Flow chromatography system (Bio-Rad).

Techniques: Recombinant, Chromatography, Incubation, Activity Assay, SDS Page, Staining