Samples were incubated for 30 min a

Samples were incubated for 30 min at 4 ◦C and centrifuged at 14,000 rpm and 4 ◦C for another 30 min. The supernatant and polymer blocks were removed and the protein pelletre-dissolved in 10 µL lysis buffer (50mM Tris/HCl pH 8.0,2mM dithiothreitol,10% v/v glycerol,0.5% v/v Tween 20,10% toluol).For protein precipitation with ethanol, 50 µL SF or thePE-UHMW blocks were mixed with 450 µL precooled ethanol (4 ◦C) and incubated at −70 ◦C for 2h before centrifugation at 14,000 rpm and roomtemperature for 30 min.The polymer blocks and the supernatant were removed and ethanol residues were evaporated in the vacuum centrifuge.After drying the pellet was re-dissolved in lysis buffer.Protein concentration was determined according to Bradford. For SDS PAGE analysis protein pellets were dissolved in lithium dodecyl sulfate sample buffer (26.5mMTris–HCl, 35.25mM TrisBase,0.5% lithium dodecyl sulfate,2.5% glycerol, 0.1275mM EDTA,0.055mM SERVA Blue G250,0.044mM Phenol Red, pH8.5,50mM dithiothreitol) at 95 ◦C for 5 min and applied to aprecast NuPAGE4–12% Bis-Tris polyacrylamide gel (Invitrogen,USA).Electrophoresis was conducted at 125V(60mA max., 12.5W)in a XcellSurelock Mini Cell electrophoresis system (Invitrogen,USA).Gels were silver stained for protein detection.