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Histone demethylase that specifically demethylates 'Lys-4' of histone H3, thereby playing a central role in histone code.
Our Abpromise guarantee covers the use of ab78322 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use 1µg for 106 cells. ab170191-Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.|
|ICC/IF||Use a concentration of 1 - 10 µg/ml.|
|WB||Use a concentration of 1 µg/ml. Predicted molecular weight: 196 kDa.|
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: KDM5A 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 - ab78322 observed at 240 kDa. Red - loading control, ab176560, observed at 50 kDa.
ab78322 was shown to specifically react with KDM5A in wild type HAP1 cells along with additional cross reactive bands. No bands were observed when KDM5A knockout samples were examined. Ab78322 and ab176560 (Rabbit anti alpha Tubulin loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10000 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.
ab78322 staining KDM5A / Jarid1A / RBBP2 [18E8] in HeLa cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4 % paraformaldehyde, permeabilized with Triton X-100 0.25% in PBS and blocked with 1.5% BSA for 30 minutes. Samples were incubated with primary antibody (1/200 in PBS + 1% BSA) for overnight at 4°C. An Alexa Fluor®488-conjugated Goat anti-mouse IgG polyclonal (1/1000 in PBS + 1% BSA) was used as the secondary antibody and incubated for 1 hour at room temperature. DAPI was used to stain the nuclear DNA.
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