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Our Abpromise guarantee covers the use of ab75607 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 5 µg/ml.|
|WB||Use a concentration of 5 - 10 µg/ml. Predicted molecular weight: 113 kDa.|
Lane 1: HeLa cell extract treated with vehicle (20 µg).
Lane 2: HeLa cells extract treated with staurosporine (20 µg).
Lane 3: Jurkat cell extract treated with vehicle (20 µg).
Lane 4: Jurkat cell extract treated with staurosporine (20 µg).
SDS PAGE performed under reducing conditions (100mM DTT, sample heated at 50°C).
Primary (Lanes 1-4): ab75607 at 2 µg/mL in 5% milk + PBS overnight at 4°C.
Secondary (Lanes 1-4): HRP-conjugated goat anti-rabbit IgG (H&L) at 1/10000 in 5% milk + PBS for 2 hour at room temperature.
Developed: ECL with 30 second exposure. Blocked in 5% milk + PBS for 3 hours at room temperature.
Predicted band size: 113 kDa and 89 kDa.
Observed band size: 113 kDa and 89 kDa.
ICC/IF image of ab75607 stained HEK293 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 (ab756007, 5µg/ml) overnight at +4°C. 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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"