After supplementation of 25 µl of protein sample loading buffer the samples were boiled for 5 min.
After the addition of SDS-PAGE buffer the samples were heated to 95°C and analyzed by Western blot using an anti-HA antiserum.
After 3 washes with wash buffer the sample was fixed with 2% glutaraldehyde for 5 minutes before staining with 2% uranyl acetate.
After 30 min, the controller switched off automatically and the cathode chamber buffer (the sample) was emptied into an Eppendorf vial.
After dialysis against the Tris-HCl buffer, the sample was further purified to remove endotoxin by polymyxin B column (Bio-Rad, USA).
After washing with 0.1 M phosphate buffer, the sample was dehydrated by a series of incubations in 30, 50 and 70% (v/v) ethanol.
After washing with 0.1 M phosphate buffer, the sample was dehydrated by a series of incubations in 30%, 50% and 70%, ethanol.
After filtration and washing with the phosphate buffer, the samples were utilized to immobilize HRP.
After three washes with NP-40 lysis buffer, the samples were boiled with 2× SDS loading buffer and subjected to SDS-polyacrylamide gel electrophoresis (PAGE) and Western blotting analysis.
Following washing in NP-40 lysis buffer, the samples were processed for Western blotting.
