SurA | SurA

application example

 Approximately 10 µg of total protein from Y. pseudotuberculosis    extracted with   were separated on 12 % SDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following  transfer in milk solution (10 % milk powder in TBST) for 1 h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 5 000 overnight at  4 °C with agitation. The antibody solution was decanted and the blot was washed twiced for 15 min in TBST at room temperature with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from GE Healthcare) diluted to 1: 20 000 in  for 1h at room temperature with agitation. The blots were washed as above and developed for 35 min with homemade ECL  detection reagent. Exposure time was 60  seconds.