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Synthetic peptide corresponding to residues on the N-terminus in Human HDAC2 protein (Q92769).
This product is a recombinant rabbit monoclonal antibody.
Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.
Our Abpromise guarantee covers the use of ab124974 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/10000 - 1/50000. Predicted molecular weight: 55 kDa.|
|IP||1/10 - 1/100.|
|ChIP||Use 10 µg for 25 µg of chromatin.
Please note product formulation is not optimised for ChIP application
Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: HDAC2 knockout HAP1 whole cell lysate (20 µg)
Lane 3: A431 whole cell lysate (20 µg)
Lane 4: Hela whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab124974 observed at 60 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab124974 was shown to recognize HDAC2 when HDAC2 knockout samples were used, along with additional cross-reactive bands. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE. ab124974 and ab8245 (Mouse anti GAPDH loading control) were but diluted at 1/10000 and incubated overnight at 4°C. Blots were developed with 800CW Goat anti Rabbit and 680CW Goat anti Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 10µg of ab124974 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach). Primers and probes are located in the first kb of the transcribed region.