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Our Abpromise guarantee covers the use of ab77269 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration. Preliminary experiments in Human Pancreas lysates gave no specific signal but low background (at antibody concentration up to 1µg/ml).|
|IHC-P||Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|ELISA||Use at an assay dependent concentration. Antibody detection limit for peptide ELISA 1:4000|
|ICC/IF||Use a concentration of 5 µg/ml.|
ICC/IF image of ab77269 stained CaCo2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal donkey 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 ab77269 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 donkey anti- goat (ab96931) 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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