Cell signaling technology (CST)
P21 Waf1/Cip1 Rabbit mAb
#2947
12D1
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Staining of p21 in human Hela S3 cells, good signal overall
Western Blot
Hela cells, cell lysate
1:1000 milk 5% in PBS 0,05% tween
Milk 5% in PBS 0,05% tween
Rabbit 1:3000 milk 5% in PBS 0,05% tween 1 hour
None
ECL
The signal is very strong when used following the CST datasheet instructions. Hela cells WCL has been loaded on a 4-12% SDS-precast gel (1:15h; 180 V) then proteins have been transferred on nitrocellulose membrane using the semi-dry blotting (1h 11V). Blocking with milk 5% in PBS 0,05% tween (1 hour) was followed by incubation overnight (1:1000 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 1-hour incubation with Rabbit secondary antibody 1:3000 in milk 5% in PBS 0,05% tween.To wash out the secondary antibody, a second extensive wash with PBS 0,05% tween (3x 5 minutes) was performed. ECL substrate has been used to detect the signal.
Very clean
It can have some specifics.
Recommended.