Synthetic peptide (Human)
This product is a recombinant rabbit monoclonal 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
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 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 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.