Overview

  • Product name
    Anti-ERG antibody [EPR3864(2)]
    See all ERG primary antibodies
  • Description
    Rabbit monoclonal [EPR3864(2)] to ERG
  • Host species
    Rabbit
  • Specificity
    This antibody does not cross-react with Fli1.
  • Tested applications
    Suitable for: ICC/IF, WB, IP, IHC-P, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide within Human ERG aa 450 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: P11308

  • Positive control
    • Jurkat whole cell lysate (ab7899), 293T cell lysate, MCF7 cell lysate, ERG recombinant protein, Human prostate adenocarcinoma tissue.
  • 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab133264 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 1/500.
WB 1/2000. Detects a band of approximately 55 kDa (predicted molecular weight: 54 kDa).
IP 1/30.
IHC-P 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Flow Cyt 1/120.

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

  • All lanes : Anti-ERG antibody [EPR3864(2)] (ab133264) at 1/2000 dilution (purified)

    Lane 1 : Jurkat cell lysate
    Lane 2 : HEK293 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 54 kDa
    Observed band size: 55 kDa
    why is the actual band size different from the predicted?



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • ab133264 staining ERG in the human cell line MCF-7(human breast carcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde, permeabilized with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/210. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

  • Immunohistochemical staining of paraffin embedded human colonic carcinoma with purified ab133264 at a working dilution of 1 in 250. The secondary antibody used is a HRP goat anti-rabbit H+L (ab97051). The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Immunohistochemical staining of paraffin embedded mouse cardiac muscle with purified ab133264 at a working dilution of 1 in 250. The secondary antibody used is a HRP goat anti-rabbit H+L (ab97051). The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Immunofluorescence staining of MCF7 cells with purified ab133264 at a working dilution of 1 in 500, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti rabbit (ab150077), used at a dilution of 1 in 500. ab7291 was used to stain tubulin, and this is shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative control is shown in bottom middle and right hand panels - for the negative controls, purified ab133264 was used at a dilution of 1/200 followed by an Alexa Fluor® 594 goat anti-mouse antibody at a dilution of 1/500.

  • ab133264 (purified) at 1/30 immunoprecipitating ERG in HEK293 cells (Lane 1). Lane 2 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-ERG antibody [EPR3864(2)] (ab133264) at 1/1000 dilution (unpurified)

    Lane 1 : Jurkat cell lysate
    Lane 2 : 293T cell lysate
    Lane 3 : MCF7 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution

    Predicted band size: 54 kDa

  • All lanes : Anti-ERG antibody [EPR3864(2)] (ab133264) at 1/1000 dilution (unpurified)

    Lane 1 : Fli1 recombinant protein
    Lane 2 : ERG recombinant protein

    Lysates/proteins at 0.01 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution

    Predicted band size: 54 kDa

  • Immunohistochemical analysis of paraffin embedded human prostate adenocarcinoma tissue labelling ERG with ab133264 at  a dilution of 1/100.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References

This product has been referenced in:
  • Cao Z  et al. The preoperative neutrophil-to-lymphocyte ratio is not a marker of prostate cancer characteristics but is an independent predictor of biochemical recurrence in patients receiving radical prostatectomy. Cancer Med 8:1004-1012 (2019). Read more (PubMed: 30693666) »
  • Yamamoto R  et al. Selective dissociation between LSD1 and GFI1B by a LSD1 inhibitor NCD38 induces the activation of ERG super-enhancer in erythroleukemia cells. Oncotarget 9:21007-21021 (2018). Read more (PubMed: 29765516) »
See all 15 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: cell conditioner 1
Permeabilization
No
Specification
lung
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Apr 22 2019

Application
ChIP
Sample
Human Cell lysate - nuclear (KG1 leukemic cell line)
Negative control
Negative control region near the AFP gene. IP using IgG
Specification
KG1 leukemic cell line
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% formaldehyde in PBS, room t
Positive control
3 positive control regions were used - the ERG +85 enhancer (ERG binding to this region in KG1 cells was published in Diffner et al Blood 2013 PMID:23327922) - plus two additional unpublished regions. IP was done in parallel with the sc354X antibody used in Diffner et al.

Dr. Julie Thoms

Verified customer

Submitted Feb 28 2017

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 28°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate
Sample
Mouse Tissue sections (kidney)
Specification
kidney
Permeabilization
Yes - 0,5% for 25 min
Fixative
Paraformaldehyde

Herr Dr. Jan Sradnick

Verified customer

Submitted Apr 08 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 30 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 28°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate
Sample
Mouse Tissue sections (kidney)
Specification
kidney
Permeabilization
Yes - 0.5% for 25 min
Fixative
zinc

Herr Dr. Jan Sradnick

Verified customer

Submitted Apr 08 2015

Answer

We looked into this about 3 years ago when one of our customers reported that ab110639 and ab92513 would recognize Fli-1. Her data demonstrated that immunoprecipitation with ab92513 and mass-spec sequencing from the CMK cell line immunoprecipitates the Fli-1 protein.
After receiving this feedback, we performed further testing to help make a final conclusion with regards to these two antibodies. After verifying the antibodies against both ERG and Fli-1 recombinant proteins the data showed that these antibodies are capable of recognizing both ERG and Fli-1. ab133264 was sub-cloned after the cross-reactivity was determined. Prior to releasing ab133264, we tested the antibody to make sure that it solely recognizes ERG. The ab92513 is from a single clone but it shares homology with Fli-1. Although ab92513 and ab133264 are generated against the same peptide, given the length of the immunogen peptide, it is possible that the antibody generated from the same peptide can recognize different epitopes of target protein.

In short, we would guarantee that ab133264 does not cross-react with Fli1.

Read More

Answer


I am happy to provide you with the concentration in this case: 2mg/ml.


.

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Question
Answer

Unpurified antibodies will not have a concentration stated on the datasheet. For most whole antiserum, culture supernatant or ascites fluid products the concentration has not been determined. Unpurified antibody preparations vary significantly in specific antibody concentration. Please be aware that this antibody is culture supernatant.

However, for some antibodies we can provide the concentration even when they are culture supernatant or ascites.

To check this, we would need the lot# of the antibody you bought from us.


Read More

Answer

Thank you for contacting us.



While the antibodies were raised against the same sequences, they may detect different epitopes. They are also different in terms of species reactivity and the applications they have been validated for. Ab133264 is recommended for use with human samples and tested positive in WB,IP,IHC and ICC, whereas ab92513 is recommended for use with mouse, rat and human and has tested positive in WB,IHC and ICC.


I hope this helps. Please do not hesitate to contact us for further assistance.

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