Overview

  • Product name

    Anti-ERG antibody [EPR3864] (Alexa Fluor® 647)
    See all ERG primary antibodies
  • Description

    Rabbit monoclonal [EPR3864] to ERG (Alexa Fluor® 647)
  • Host species

    Rabbit
  • Conjugation

    Alexa Fluor® 647. Ex: 652nm, Em: 668nm
  • Tested applications

    Suitable for: ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human ERG aa 450 to the C-terminus.
    Database link: P11308

  • Positive control

    • ICC/IF: THP-1 cells
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab196149 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/100.

This product gave a positive signal in THP-1 cells fixed with 4% formaldehyde (10 min)

Flow Cyt Use at an assay dependent concentration.

Target

  • Function

    Transcriptional regulator. May participate in transcriptional regulation through the recruitment of SETDB1 histone methyltransferase and subsequent modification of local chromatin structure.
  • Involvement in disease

    Defects in ERG are a cause of Ewing sarcoma (ES) [MIM:612219]. A highly malignant, metastatic, primitive small round cell tumor of bone and soft tissue that affects children and adolescents. It belongs to the Ewing sarcoma family of tumors, a group of morphologically heterogeneous neoplasms that share the same cytogenetic features. They are considered neural tumors derived from cells of the neural crest. Ewing sarcoma represents the less differentiated form of the tumors. Note=A chromosomal aberration involving ERG is found in patients with Erwing sarcoma. Translocation t(21;22)(q22;q12) with EWSR1.
    Note=Chromosomal aberrations involving ERG have been found in acute myeloid leukemia (AML). Translocation t(16;21)(p11;q22) with FUS. Translocation t(X;21)(q25-26;q22) with ELF4.
  • Sequence similarities

    Belongs to the ETS family.
    Contains 1 ETS DNA-binding domain.
    Contains 1 PNT (pointed) domain.
  • Cellular localization

    Nucleus. Cytoplasm. Localized in cytoplasmic mRNP granules containing untranslated mRNAs.
  • Information by UniProt
  • Database links

  • Alternative names

    • Avian erythroblastosis virus E-26 (v-ets) oncogene related antibody
    • D030036I24Rik antibody
    • Erg 3 antibody
    • Erg antibody
    • ERG/EWS fusion gene, included antibody
    • ERG/FUS fusion gene, included antibody
    • ERG/TMPSSR2 fusion gene, included antibody
    • ERG_HUMAN antibody
    • ERG1, included antibody
    • ERG2, included antibody
    • ets related antibody
    • ETS-related gene antibody
    • KCNH2 antibody
    • Oncogene ERG antibody
    • p55 antibody
    • TMPRSS2/ERG fusion antibody
    • transcriptional regulator ERG (transforming protein ERG) antibody
    • Transcriptional regulator ERG antibody
    • Transforming protein ERG antibody
    • v ets avian erythroblastosis virus E26 oncogene antibody
    • v ets avian erythroblastosis virus E26 oncogene related antibody
    • v ets erythroblastosis virus E26 oncogene homolog antibody
    • v ets erythroblastosis virus E26 oncogene like antibody
    • v ets erythroblastosis virus E26 oncogene like isoform 2 antibody
    • v-ets erythroblastosis virus E26 oncogene antibody
    • v-ets erythroblastosis virus E26 oncogene homolog (avian) antibody
    • V-ets erythroblastosis virus E26 oncogene like (Avian), isoform CRA_e antibody
    see all

Images

  • For mouse retina immunostaining, eyes were collected at the indicated time points and fixed in 4% PFA in PBS for 1 h at room temperature (RT). After two PBS washes, retinas were micro-dissected and stained. Briefly, retinas were blocked and permeabilized with 0.3% Triton X-100, 3% fetal bovine serum (FBS) and 3% donkey serum in PBS. Samples were then washed twice in PBLEC buffer (1 mM CaCl2, 1 mM MgCl2, 1 mM MnCl2 and 1% Triton X-100 in PBS). Biotinylated isolectinB4 or primary antibodies (Panel a, ab196149 1/100 dilution) were diluted in PBLEC buffer and tissues were incubated in this solution for 2 h at RT or overnight at 4 °C. After five washes in blocking solution diluted 1:2, samples were incubated for 1 h at RT with Alexa-conjugated secondary antibody. After two washes in PBS, retinas were mounted with Fluoromount-G. To detect EdU-labeled DNA, an additional step was performed before mounting using the Click-It EdU kit.

    Dll4/Notch signalling inhibition induces context-dependent proliferative effects. a–d Confocal micrographs of the postnatal retinal vasculature from animals treated at P5 with IgG (control) or anti-Dll4 for 24 h (a, b) or 48 h (c, d). Anti-Erg (red) labels EC nuclei. EdU labels the nuclei of all cells in S-phase in the previous 4 h. Blue nuclei mark non-endothelial cells in S-phase, and double-positive (Erg+/EdU+) cell nuclei are pseudocoloured green to better highlight ECs in S-phase. Scale bars, 200 μm.

     

  • RLCs selectively differentiate to intraglomerular mesangial cells during EC model.
    Panel A shown only. Representative confocal microscopy images for day 0 and day 7 of β-gal and the EC markers ERG (upper panels) or CD31 (lower panels) co-stained kidney slices.

    Immunostainings and Acid fuchsin orange G (AFOG) staining were performed on paraffin-embedded 4-μm kidney sections. Antibodies were diluted in 1% BSA/TBS. Primary antibodies were incubated overnight at 4°C, secondary antibodies and Avidin D-conjugated horseradish peroxidase were incubated for 2 hours at room temperature. All immunofluorescence samples were counterstained with the nuclear marker 4′,6-diamidino-2-phenylindole (DAPI) and mounted with mowiol mounting medium. Endogenous peroxidase activity was suppressed with 3% hydrogen peroxide solution. Samples were counterstained with hematoxylin, dehydrated and mounted. The following antibodies were used: WT-1 (ab202639), nephrin, PDGFRβ (ab91066), α8-integrin, CD31, ERG (ab196149), β-galactosidase (ab9361), renin, CD45 (ab64100).

  • Mt4-mmp expression during early mouse embryonic development.


    Panel K shown only: Double-labeled cells for the endothelial markers ERG (red, arrows in K) and β-gal (green) demonstrate that cells expressing Mt4-mmp in this location are endothelial cells. Abbreviations: da, dorsal aorta; fp, floor plate; s, somite; NCC, neural crest cells; NT, neural tube. Scale bars: 30 μm (J-L).

    For immunohistochemical procedures, embryos were fixed in 4% PFA in PBS 0.1M pH 7.2 by immersion or perfusion depending on the developmental stage, cryoprotected in a 30% sucrose solution, and then embedded in OCT and sectioned in the cryostat at 20 μm thickness in the transverse plane. Immunohistochemistry was performed following standard protocols. Primary antibodies used include: polyclonal anti-CD31 hamster (1/1000), anti-β-galactosidase rabbit (1/1000; ab4761, Abcam), anti-FoxA2 mouse (1/250), anti-Nkx6.1 mouse (1/1000), anti-Olig2 rabbit (1/1000), anti-ERG-647 rabbit (1/500; ab196149, Abcam) and anti-WT-1 mouse (Wilms Tumor-1; 1/50;). Sections were incubated with the primary antibody diluted in PBS containing 0.1% Triton X-100 and 1% bovine serum albumin (BSA), for 48 h at 4°C. Subsequently, the sections were rinsed in PBS and incubated for 2 hours at room temperature with 488 or 594-Alexa™-conjugated fluorescent antibodies (1/1000). Sections were counterstained with Hoechst (1:1000) for 5 min at room temperature to visualize nuclei.

  • ab196149 staining ERG in THP-1 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab196149 at 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Flow Cytometry analysis of A431(human epidermoid carcinoma) cells labeling ERG with purified ab196149 at 1/500 dilution(10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 647) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

References

This product has been referenced in:

  • Pontes-Quero S  et al. High mitogenic stimulation arrests angiogenesis. Nat Commun 10:2016 (2019). IHC (PFA fixed) ; Mouse . Read more (PubMed: 31043605) »
  • Fernández-Chacón M  et al. iSuRe-Cre is a genetic tool to reliably induce and report Cre-dependent genetic modifications. Nat Commun 10:2262 (2019). IHC (PFA fixed) ; Mouse . Read more (PubMed: 31118412) »
See all 4 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Peripheral lymph node)
Permeabilization
Yes - 0.05% Tween, 0.1% Triton
Specification
Peripheral lymph node
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Fixative
Paraformaldehyde

Dr. Agata Szade

Verified customer

Submitted Dec 06 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Kidney)
Antigen retrieval step
None
Permeabilization
Yes - 0.5% Triton-X
Specification
Kidney
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 26°C
Fixative
Zink

Abcam user community

Verified customer

Submitted Jun 17 2016

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