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Recombinant Rat ATPase Inhibitory Factor 1 (UniProtKB ID: Q6IAQ7).
Our Abpromise guarantee covers the use of ab110277 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 10 µg/ml.|
|Flow Cyt||Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 10,18 kDa (predicted molecular weight: 12 kDa).|
|ELISA||Use a concentration of 4 µg/ml. (In-Cell ELISA, 0.4 µg/well).|
Lanes 1 - 3: Merged signal (red and green). Green - ab110277 observed at 12 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab110277 was shown to recognize ATPase Inhibitory Factor 1 in wild-type HAP1 cells as signal was lost at the expected MW in ATPIF1 (ATPase Inhibitory Factor 1) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and ATPIF1 (ATPase Inhibitory Factor 1) knockout samples were subjected to SDS-PAGE. Ab110277 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
ICC/IF image of ab110277 stained HepG2 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 (ab110277, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse 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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