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An ocular melanoma cell line expressing high levels of human integrin alpha V beta 1.
Our Abpromise guarantee covers the use of ab11470 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use at an assay dependent concentration.|
|ICC/IF||Use at an assay dependent concentration.|
|WB||Use at an assay dependent concentration.|
|Flow Cyt||Use a concentration of 1 - 10 µg/ml. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.|
ICC/IF image of ab11470 stained A549 cells. The cells were 4% formaldehyde 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 (ab11470, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, a goat anti-mouse DyLight® 488 (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.
Overlay histogram showing A549 cells stained with ab11470 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab11470, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was a goat anti-mouse DyLight® 488 (IgG; H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive result in 4% paraformaldehyde (10 min) fixed A549 cells used under the same conditions.
Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.