Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR12021] to IMP3
- Suitable for: WB, ICC/IF, IP, Flow Cyt
- Knockout validated
- Reacts with: Mouse, Rat, Human
Product nameAnti-IMP3 antibody [EPR12021]
See all IMP3 primary antibodies
DescriptionRabbit monoclonal [EPR12021] to IMP3
Tested applicationsSuitable for: WB, ICC/IF, IP, Flow Cytmore details
Unsuitable for: IHC-P
Species reactivityReacts with: Mouse, Rat, Human
Recombinant fragment within Human IMP3 aa 1-250. The exact sequence is proprietary.
Database link: O00425
- HeLa, BxPC-3 and 293T lysates; BxPC-3 cells.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
Concentration information loading...
PurityTissue culture supernatant
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.|
FunctionRNA-binding protein that act as a regulator of mRNA translation and stability. Binds to the 5'-UTR of the insulin-like growth factor 2 (IGF2) mRNAs. Binds to sequences in the 3'-UTR of CD44 mRNA.
Tissue specificityExpressed in fetal liver, fetal lung, fetal kidney, fetal thymus, fetal placenta, fetal follicles of ovary and gonocytes of testis, growing oocytes, spermatogonia and semen (at protein level). Expressed in cervix adenocarcinoma, in testicular, pancreatic and renal-cell carcinomas (at protein level). Expressed ubiquitously during fetal development at 8 and 14 weeks of gestation. Expressed in ovary, testis, brain, placenta, pancreatic cancer tissues and pancreatic cancer cell lines.
Sequence similaritiesBelongs to the RRM IMP/VICKZ family.
Contains 4 KH domains.
Contains 2 RRM (RNA recognition motif) domains.
DomainThe third and fourth KH domains are important for binding to the untranslated region (UTR) of target mRNA.
Cellular localizationNucleus. Cytoplasm. Found in lamellipodia of the leading edge, in the perinuclear region, and beneath the plasma membrane. The subcytoplasmic localization is cell specific and regulated by cell contact and growth. Localized at the connecting piece and the tail of the spermatozoa. Colocalized with CD44 mRNA in RNP granules.
- Information by UniProt
- Cancer/testis antigen 98 antibody
- CT98 antibody
- DKFZp686F1078 antibody
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.
All lanes : Anti-IMP3 antibody [EPR12021] (ab177477) at 1/1000 dilution
Lane 1 : HeLa lysate
Lane 2 : BxPC-3 lysate
Lane 3 : 293T lysate
Lysates/proteins at 10 µg per lane.
All lanes : HRP labeled goat anti-rabbit at 1/2000 dilution
Predicted band size: 63 kDa
Immunoprecipitation of IMP3 from 293T cell lysate using ab177477 at 1/10.
All lanes : Anti-IMP3 antibody [EPR12021] (ab177477) at 1/20000 dilution
Lane 1 : MDA-MB-231 (human breast adenocarcinoma) whole cell lysate
Lane 2 : MDA-MB-435S (human melanoma) whole cell lysate
Lane 3 : MCF-7 (human breast adenocarcinoma) whole cell lysate
Lane 4 : Human Fetal Kidney
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) at 1/2000 dilution
Predicted band size: 63 kDa
Observed band size: 69 kDa why is the actual band size different from the predicted?
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
ab177477 has been referenced in 3 publications.
- Legrand JMD et al. DDX5 plays essential transcriptional and post-transcriptional roles in the maintenance and function of spermatogonia. Nat Commun 10:2278 (2019). PubMed: 31123254
- You S et al. Epithelial-mesenchymal transition in colorectal carcinoma cells is mediated by DEK/IMP3. Mol Med Rep 17:1065-1070 (2018). PubMed: 29115492
- Lu L et al. The expression of IMP3 in 366 cases with ovarian carcinoma of high grade serous, endometrioid and clear cell subtypes. Pathol Res Pract N/A:N/A (2018). IHC-P . PubMed: 29866423