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Purified human beta 1 preparation from HT1080 fibrosarcoma cell extract.
ab30483 is identified as capable of immunoprecipitating a non-reducible alpha subunit from I125 surface labelled Human cell line extract. It is confirmed as specific to the alpha 2 subunit by relative expression of antigen on various cell lines by FACS, and its recognition of affinity purified alpha 2 beta 1 in dot blots.
Our Abpromise guarantee covers the use of ab30483 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.|
|IP||Use a concentration of 25 - 50 µg/ml.|
|Flow Cyt||Use 0.1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.|
|ELISA||Use a concentration of 10 µg/ml.|
|IHC-Fr||Use a concentration of 30 - 40 µg/ml.|
Overlay histogram showing HT1080 cells stained with ab30483 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab30483, 0.1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was a goat anti-mouse Alexa Fluor® 488 (IgG; H+L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HT1080 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
ab30483 staining human breast cancer cells by ICC/IF. Cells were PFA fixed and blocked with 1% serum for 16 hours at 20°C prior to incubating with ab30483 (at 30µg/ml) for 16 hours at 4°C. An Alexa Fluor® 488 conjugated goat anti-mouse antibody, diluted 1/200, was used as the secondary.