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Our Abpromise guarantee covers the use of ab81149 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 22, 34, 64 kDa (predicted molecular weight: 64 kDa). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.|
|ELISA||Use at an assay dependent concentration.
ELISA titre using peptide based assay 1/62500.
|IHC-P||Use a concentration of 5 µg/ml.|
|ICC/IF||Use a concentration of 5 µg/ml.|
ab81149 stained HeLa cells. The cells were 4% formaldehyde fixed for 10 minutes at room temperature and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab81149 at 5µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate tissue labelling ARID5A with ab81149 at 5.0µg/ml.
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