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Synthetic peptide corresponding to residues in Human MBD3 (UniProt O95983).
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents
This product is a recombinant rabbit monoclonal antibody.
Our Abpromise guarantee covers the use of ab157464 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000 - 1/5000. Predicted molecular weight: 33 kDa.|
|ICC/IF||1/100 - 1/1000.|
|Flow Cyt||1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|IP||Use at an assay dependent concentration.|
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: ProteinX knockout HAP1 cell lysate (20 µg)
Lane 3: Wild-type HAP1 cell lysate (20 µg)
Lane 4: ProteinX knockout HAP1 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab157464 observed at 33 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab157464 was shown to recognize MBD3 when MBD3 knockout samples were used, along with additional cross-reactive bands. Wild-type and MBD3 knockout samples were subjected to SDS-PAGE. ab157464 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) labelling MBD3 with purified ab157464 at 1/1000. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).
Control: PBS only
Immunoprecipitation of MBD3, using WT and MBD3-/- ES cells (however this was not a 100% pure MBD3 null population). The protein was immunoprecipitated using 1000µg nuclear extract, and 1.03µg of Rabbit polyclonal to MBD3 (ab157464). The same MBD3 antibody was used for the western blot. Left blot image taken after a short exposure, right image taken after a longer exposure.
Lanes 1, 3 and 5: Mbd3+/+, Lanes 2, 4 and 6: Mbd3-/- (not 100% pure Mbd3 null population), Lanes 1 and 2: Input, Lanes 3 and 4: IP with ab91458, Lanes 5 and 6: IP with ab157464.
Band: 33kDa: MBD3
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"