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A synthetic peptide corresponding to Human USP13.
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 ab109264 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 - 1/10000. Detects a band of approximately 97 kDa (predicted molecular weight: 97 kDa).|
|IP||1/10 - 1/100.|
|ICC||1/100 - 1/250.|
|Flow Cyt||1/940 - 1/9400. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.|
|ICC/IF||Use a concentration of 10 µg/ml.|
Immunocytochemistry/Immunofluorescence analysis of Neuro-2a (Mouse neuroblastoma cell line) labeling USP13 with Purified ab109264 at 1/500 dilution (5 µg/ml). Cells were fixed with 100% methanol. ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) at 1/1000 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: USP13 knockout HAP1 cell lysate (20 µg)
Lane 3: A375 cell lysate (20 µg)
Lane 4: SH-SY5Y cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab109264 observed at 100 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab109264 was shown to recognize USP13 when USP13 knockout samples were used, along with additional cross-reactive bands. Wild-type and USP13 knockout samples were subjected to SDS-PAGE. ab109264 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 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.
ICC/IF image of ab109264 stained MCF-7 cells. The cells were 100% methanol fixed (5 min) 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 ab109264 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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.
ab109264 has not yet been referenced specifically in any publications.