Roche
GFP antibody clones 7.1 and 13.1
11814460001
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GFP antibody has been used to detect the GFP signal after the transfection of an empty-GFP plasmid in Hela cells.
Western Blot
Hela cells, cell lysate
1:2000 Milk 5% in PBS 0,05% tween 1h RT
Milk 5% in PBS 0,05% tween
Mouse 1:3000 milk 5% in PBS 0,05% tween 45 minutes
None
ECL
Hela cells WCL has been loaded on a 4-12% SDS-precast gel (1:30h; 180 V) then proteins have been transferred on nitrocellulose membrane using the semi-dry blotting (1h 12V). Blocking with milk 5% in PBS 0,05% tween (45 minutes) was followed by incubation overnight at room temperature with anti- GFP antibody (1:2000 in milk 5% in PBS 0,05% tween). Then the membrane was washed with 3x 5 minutes with PBS 0,05% tween, followed by a 45 minutes incubation with Mouse secondary antibody 1:3000 in milk 5% in PBS 0,05% tween. The secondary antibody was washed out with a second wash with PBS 0,05% tween (3x 5 minutes). ECL substrate has been used to detect the signal.
Very good staining and results obtained.
Sometimes the signal can be very strong, the optimization of the dilution of the antibody between 1:2000-5000 can helo
Great antibody