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Recombinant fragment within Human IMP3. The exact sequence is proprietary.
Database link: O00425
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 ab177477 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 - 1/5000. Detects a band of approximately 69 kDa (predicted molecular weight: 63 kDa).|
|ICC/IF||1/50 - 1/100.|
|IP||1/10 - 1/100.|
|Flow Cyt||Use at an assay dependent concentration.|
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: IGF2BP3 (IMP3) knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: HEK293 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab177477 observed at 70 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab177477 was shown to specifically react with IGF2BP3 (IMP3) in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when IGF2BP3 (IMP3) knockout cells were examined. Wild-type and IGF2BP3 (IMP3) knockout samples were subjected to SDS-PAGE. Ab177477 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1/20,000 dilution for 1 hour at room temperature before imaging.
Immunofluorescent analysis of BxPC-3 cells labeling IMP3 with ab177477 at 1/50 (green) and DAPI staining (blue).
Flow cytometry analysis of BxPC-3 (human pancreas adenocarcinoma) cells labeling with purified ab177477 at 1/300 dilution ( 10ug/ml) (Red). Cells were fixed with 80% methanol and permeabilised with 0.1% Tween-20. A Goat anti rabbit IgG (Alexa Fluor® 488)(ab150077) )(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black)(ab172730) was used as a isotype control.Cell without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.
Immunoprecipitation of IMP3 from 293T cell lysate using ab177477 at 1/10.
Blocking/Diluting buffer 5% NFDM/TBST
The protein expression of IMP-3 in MCF-7 cells is negative or undetectable (PMID: 22266872).
Lane 1: 10 seconds
Lane 2-4: 5 seconds
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"