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Our Abpromise guarantee covers the use of ab12171 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ELISA||Use at an assay dependent dilution.|
|ICC||Use at an assay dependent dilution.|
|IP||Use at an assay dependent dilution.|
|WB||Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 140 kDa (predicted molecular weight: 119 kDa).|
|ChIP||Use at an assay dependent concentration. PubMed: 21586557|
Lane 1: Wild-type HAP1 cell lysate (40 µg)
Lane 2: HDAC4 knockout HAP1 cell lysate (40 µg)
Lane 3: HeLa cell lysate (40 µg)
Lane 4: NIH3T3 cell lysate (40 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab12171 observed at 140 kDa. Red - loading control, ab176560, observed at 52 kDa.
ab12171 was shown to recognize HDAC4 when HDAC4 knockout samples were used, along with additional cross-reactive bands. Wild-type and HDAC4 knockout samples were subjected to SDS-PAGE. Ab12171 and ab176560 (loading control to alpha tubulin) were diluted at 1 µg/ml and 1:10,000 dilution respectively and incubated overnight at 4C. 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.
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