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Our Abpromise guarantee covers the use of ab57632 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 - 5 µg/ml. This antibody has only been tested in WB against the recombinant fragment used as immunogen. We have no data on the detection of endogenous protein.|
|Flow Cyt||Use 0.5µg for 106 cells.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: VPS35 knockout HAP1 cell lysate (20 µg)
Lane 3: NIH/3T3 cell lysate (20 µg)
Lane 4: A549 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab57632 observed at 91 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab57632 was shown to specifically react with VPS35 in wild-type HAP1 cells. No band was observed when VPS35 knockout samples were examined. Wild-type and VPS35 knockout samples were subjected to SDS-PAGE. ab5732 and ab181602 (loading control to GAPDH) were diluted 1µg/mL and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.