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Our Abpromise guarantee covers the use of ab36938 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 µg/ml. Predicted molecular weight: 154 kDa.Can be blocked with Human KDM6A / UTX peptide (ab36937).|
|ICC/IF||Use a concentration of 1 µg/ml.|
|ChIP||Use at an assay dependent concentration. PubMed: 26306033|
Lanes 1 & 3: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2 & 4: KDM6A knockout HAP1 whole cell lysate (20 µg)
Lanes 1 - 2: Green - ab36938 observed at 154 kDa.
Lanes 3 - 4: Red - loading control, ab18058, observed at 130 kDa.
ab36938 was shown to recognize KDM6A in wild-type HAP1 cells as signal was lost at the expected MW in KDM6A knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and KDM6A knockout samples were subjected to SDS-PAGE. Ab36938 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 0.7 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Lane 3 is NIH3T3 cells transduced with lentivirus expressing shRNA for mouse Utx.
5% milk was used as the blocking agent (incubated for 1 hour at 25°C).
ab36938 was diluted with TBS+0.1% Tween 20 and incubated for 1 hour at 25°C.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"