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Synthetic peptide. within Human MEK2 aa 1-100 (N terminal). The exact sequence is proprietary.
Database link: P36507
A trial size is available to purchase for this antibody.
Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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 ab32517 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 1 µg/ml.|
|WB||1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 44 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: MEK2 knockout HAP1 cell lysate (20 µg)
Lane 3: Jurkat cell lysate (20 µg)
Lane 4: K562 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab32517 observed at 44 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab32517 was shown to specifically react with MEK2 when MEK2 knockout samples were used. Wild-type and MEK2 knockout samples were subjected to SDS-PAGE. ab32517 and ab8245 (loading control to GAPDH) were diluted 1/10 000 and 1/2000 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 1/10 000 dilution for 1 h at room temperature before imaging.
ab32517 staining MEK2 in wild-type HAP1 cells (top panel) and MEK2 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab32517 at 1μg/ml and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab32517 overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) at a 1:10000 dilution for 1hr at room temperature and then imaged.
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