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Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human AMPK beta 1 aa 150-250. The exact sequence is proprietary.
Database link: Q9Y478
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 ab32112 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: 30 kDa.|
|IHC-P||Use at an assay dependent concentration.|
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: AMPK beta 1 knockout HAP1 cell lysate
Lane 3: HeLa cell lysate
Lane 4: A431 cell lysate
Lanes 1 - 4: Merged signal (red and green). Green - ab32112 observed at 38 kDa. Red - loading control, ab18058, observed at 124 kDa.
ab32112 was shown to specifically react with AMPK beta 1 when AMPK beta 1 knockout samples were used. Wild-type and AMPK beta 1 knockout samples were subjected to SDS-PAGE. ab32112 and ab18058 (loading control to Vinculin) were both diluted 1/10000 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 MCF-7 cells labelling AMPK beta 1 with purified ab32112 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (ab150077) at 1/1000 dilution was used as the secondary antibody. Nuclei counterstained with DAPI (blue).
Secondary Only Control: PBS was used instead of the primary antibody as the negative control.