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Our Abpromise guarantee covers the use of ab15270 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.|
|WB||Use at an assay dependent concentration. Predicted molecular weight: 312 kDa.|
|IP||Use at an assay dependent concentration.|
|IHC-P||1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
ICC/IF image of ab15270 stained A431 cells. The cells were 4% paraformalehyde fixed (10 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 (ab15270, 1µg/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.
Normal human colon section stained with ab15270 (1:100 for 10 min at RT). Sections were pre-treated by boiling in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at RT for 20 min.
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