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.
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, 0.05% BSA, 50% Tissue culture supernatant
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
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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
This product has been referenced in:
- Legrand JMD et al. DDX5 plays essential transcriptional and post-transcriptional roles in the maintenance and function of spermatogonia. Nat Commun 10:2278 (2019). Read more (PubMed: 31123254) »
- 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 . Read more (PubMed: 29866423) »