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Synthetic peptide (Human)
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.
Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Our Abpromise guarantee covers the use of ab51250 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/500. Detects a band of approximately 105 kDa (predicted molecular weight: 97 kDa).|
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|IHC-P||Use at an assay dependent concentration.|
|ICC/IF||1/100 - 1/250.|
Lane 1: Wild-type HAP1 cell lysate (40 µg)
Lane 2: Protein Kinase D2 knockout HAP1 cell lysate (40 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: HEK293 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab51250 observed at 110 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab51250 was shown to recognize Protein Kinase D2 when Protein Kinase D2 knockout samples were used, along with additional cross-reactive bands. Wild-type and Protein Kinase D2 knockout samples were subjected to SDS-PAGE. Ab51250 and ab8245 (loading control to GAPDH) were diluted at 1/500 and 1:10,000 dilution respectively and incubated overnight at 4C. Blots were developed with IRDye® 800CW Goat anti-Rabbit IgG (H + L) and IRDye® 680 Goat anti-Mouse IgG (H + L) secondary antibodies at 1:10,000 dilution for 1 hour at room temperature before imaging.
ICC/IF image of ab51250 stained HeLa cells. The cells were 4% Formaldehyde fixed (10 mins) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab51250, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.