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All lanes : Anti-BMAL1 antibody (ab93806) at 1/2000 dilution
Lane 1 : HeLa lysate at 50 µg
Lane 2 : HeLa lysate at 15 µg
Lane 3 : HeLa lysate at 5 µg
Lane 4 : 293T at 50 µg
Lane 5 : NIH3T3 at 50 µg
developed using the ECL technique
Predicted band size : 69 kDa
Exposure time : 3 minutes
Ab93806 at 1 µg/ml detecting BMAL1 in HeLa whole cell lysate by WB following IP.
Lane 1: IP with an antibody which recognizes an upstream epitope of BMAL1
Lane 2: ab93806 at 3µg/mg of lysate
Lane 3: control IgG.
In each case, 1 mg of lysate was used for IP and 20% of the IP was loaded.
Detection: Chemiluminescence an with exposure time of 30 seconds
ICC/IF image of ab93806 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab93806, 5µg/ml) overnight at +4ºC. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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