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Tissue/ cell preparation (Human): HT1080 fibrosarcoma cells.
ab24697 is effective in blocking adhesion cells to collagen coated surfaces. It immunoprecipitates the a2 subunit (reducing) or a2 complexed with ß1 from surface labeled cell lysates (non-reducing). In imunofluorescence, this antibody can be used to detect a2 in collagen dependent focal adhesions.
Our Abpromise guarantee covers the use of ab24697 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.|
|Blocking||Use at an assay dependent concentration.|
|Neutralising||Use at an assay dependent concentration.|
|Flow Cyt||Use 1µg for 106 cells. (Also PMID - 19780039)
ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|Inhibition Assay||Use at an assay dependent concentration.|
|ICC/IF||Use a concentration of 5 - 10 µg/ml.|
Overlay histogram showing HT1080 cells stained with ab24697 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab24697, 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 80% methanol (5 min) fixed HT1080 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.