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A synthetic peptide corresponding to Human USP13.
Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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.
This product is a recombinant rabbit monoclonal antibody.
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.