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Synthetic peptide conjugated to KLH derived from within residues 100 - 200 of Human Dnmt1.
(Peptide available as ab21999.)
Our Abpromise guarantee covers the use of ab19905 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. Detects a band of approximately 183 kDa (predicted molecular weight: 183 kDa).|
|ICC/IF||Use a concentration of 1 µg/ml.|
|IP||Use at an assay dependent concentration.|
|IHC-P||Use a concentration of 1 - 10 µg/ml.|
Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: DNMT1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: Hek293 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab19905 observed at 170 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab19905 was shown to recognize DNMT1 when DNMT1 knockout samples were used, along with additional cross-reactive bands. Wild-type and DNMT1 knockout samples were subjected to SDS-PAGE. Ab19905 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10000 dilution respectively. Blots were developed with 800CW Goat anti Rabbit and 680CW Goat anti Mouse secondary antibodies at 1/10010 dilution for 1 hour at room temperature before imaging.
ab19905 at a 1/200 dilution staining paraformaldehyde fixed asynchronous HeLa cells by ICC/IF. The antibody was incubated with the cells for 30 minutes and then bound antibody was detected using a Cy3 conjugated goat anti-rabbit antibody. Nuclei were visualised using DAPI staining. Ab19905 gives a pattern that is predominantly enriched within nuclei.
This image is courtesy of an Abreview submitted by Kirk McManus.
Jurkat cells were incubated at 37°C for 5 days with vehicle control (0 µM) and different concentrations of hydralazine hydrochloride (ab120863). Decreased expression of DNMT1 (ab19905) in Jurkat cells correlates with an increase in hydralazine hydrochloride concentration, as described in literature.
Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 20µg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab19905 at 1 µg/ml and ab8227 at 1 µg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.
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