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Synthetic peptide within Human AKT1 (C terminal) (phospho S473). The exact sequence is proprietary.
Our Abpromise guarantee covers the use of ab66138 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 1 µg/ml.|
|ELISA||Use a concentration of 0.01 - 0.1 µg/ml.|
|IHC-Fr||1/100 - 1/200.|
|WB||Use a concentration of 0.1 - 0.2 µg/ml. Predicted molecular weight: 55 kDa.
Blocking is recommended with BSA. Non-fat dry milk and PBS should be avoided.
|IP||Use a concentration of 2 - 5 µg/ml.|
ICC/IF image of ab66138 stained MCF-7 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 ab66138 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 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.
1µg peptide blotted onto a nitrocellulose membrane, followed by ab66138 at 1:2000 dilution. A: AKT1 (pS473) peptide B: AKT1 non-phosphopeptide C: non-related phosphopeptide