Recombinant Anti-ERG antibody [EPR3864] (ab92513)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3864] to ERG
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-ERG antibody [EPR3864]
See all ERG primary antibodies -
Description
Rabbit monoclonal [EPR3864] to ERG -
Host species
Rabbit -
Specificity
This antibody also detects Fli-1.
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Tested applications
Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Jurkat, HeLa and RAW 264.7 cell lysates; Rat brain and heart lysates. IHC-P: Human kidney, brain and prostate adenocarcinoma tissues; Fus A5 transgenic mouse prostate tissue; Mouse brain tissue. ICC/IF: Circulating tumor cells (CTCs) from a castrate-resistant prostate cancer (CRPC) patient; THP-1 cells. Flow Cyt (intra): THP-1 cells.
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General notes
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Dissociation constant (KD)
KD = 8.90 x 10 -10 M Learn more about KD -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3864 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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ChIP Related Products
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab92513 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 |
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WB | (1) |
1/1000 - 1/10000. Predicted molecular weight: 55 kDa.
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IHC-P | (3) |
1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified, use 1/100 - 1/250. |
ICC/IF | (3) |
Use a concentration of 1 µg/ml.
This product gave a positive signal in THP-1 (-ve: HCT116) fixed with 4% formaldehyde (10 min). |
Flow Cyt (Intra) |
Use at an assay dependent concentration.
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Notes |
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WB
1/1000 - 1/10000. Predicted molecular weight: 55 kDa. |
IHC-P
1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified, use 1/100 - 1/250. |
ICC/IF
Use a concentration of 1 µg/ml. This product gave a positive signal in THP-1 (-ve: HCT116) fixed with 4% formaldehyde (10 min). |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
Target
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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
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Database links
- Entrez Gene: 2078 Human
- Entrez Gene: 13876 Mouse
- Entrez Gene: 170909 Rat
- Omim: 165080 Human
- SwissProt: P11308 Human
- SwissProt: P81270 Mouse
- Unigene: 473819 Human
- Unigene: 164531 Mouse
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Alternative names
- Avian erythroblastosis virus E-26 (v-ets) oncogene related antibody
- D030036I24Rik antibody
- Erg 3 antibody
see all
Images
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Immunohistochemical analysis of formalin-fixed paraffin-embedded human kidney labelling ERG with ab92513 at a concentration of 1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32min with ULTRA cell conditioning solution (CC1 pH8.5). ab92513 anti ERG antibody was incubated at 37°C for 16min. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
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Flow cytometry overlay histogram showing left THP-1 positive cells and right negative HCT116 stained with ab92513 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab92513) (1x 106 in 100μl at 0.04μg/ml (1/54000)) for 30min at 22°C.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C
Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
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Tissue Microarrays stained for Anti-ERG antibody [EPR3864] using ab92513 in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negaive (cross mark) staining per sample type tested. The section was incubated with ab92513 for 30 mins at room temperature followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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Formalin-fixed, paraffin-embedded mouse brain tissue stained for ERG using ab92513 at 1/200 dilution in immunohistochemical analysis. A horse radish peroxidase antibody was used as the secondary antibody.
Antigen Retrieval: 40x; Proteinase K antigen retrieval - 15 min at 37 C
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All lanes : purified
Lane 1 : rat brain lysate
Lane 2 : rat heart lysate
Lane 3 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 55 kDa
Observed band size: 55 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
ab92513 staining ERG in THP-1 cells, with negative expression in HCT116 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilised 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 ab92513 at 1 μg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin at 0.5 μg/ml. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150119, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 647), pre-adsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.
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Functional characterization and detection of genetic alterations in GEDI-captured cells. The TMPRSS2:ERG fusion protein is detected in GEDI-captured circulating tumor cells (CTCs) from a castrate-resistant prostate cancer (CRPC) patient. PSMA-captured CTCs were stained on the device with ab92513. Representative examples of PSMA+/CD45− CTCs are shown, two of which are positive for ERG. Scale bars: 10 microns.
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ERG and GSTP1 immunostainings of human prostate cancer samples using ab92513.
Representative immunohistochemical images of prostate cancer samples are shown that were positive for ERG and negative for GSTP1 (A), positive for both ERG and GSTP1 (B), negative for both ERG and GSTP1(C), and negative for ERG and positive for GSTP1 (D). The internal staining control for ERG is the endothelium (arrows) and for GSTP1 the stromal and/or basal cells of normal prostate glands. N, normal prostate gland; S, Stroma; T, tumor gland. Scale bars equal 100μm
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Intracellular Flow Cytometry analysis of THP-1 (human monocytic leukemia cell line) cells labeling ERG with purified ab92513 at 1/1000 dilution (1ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluorr® 488) (ab150077) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) (ab172730) was used as the isotype control, Cell without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
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Immunohistochemical staining of paraffin embedded human kidney with purified ab92513 at a working dilution of 1/1000. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. 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.
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All lanes : Anti-ERG antibody [EPR3864] (ab92513) at 1/2000 dilution (purified)
Lane 1 : Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate
Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 55 kDa
Observed band size: 55 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
Immunofluorescence staining of THP-1 (human monocytic leukemia cell line) cells with purified ab92513 at a working dilution of 1/100, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab92513 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
Alexa Fluor® 488 (ab196374) and Alexa Fluor® 647 (ab196149) conjugated versions are available for this clone.
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Anti-ERG antibody [EPR3864] (ab92513) at 1/1000 dilution (unpurified) + Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate at 10 µg
Secondary
HRP labelled Goat anti-Rabbit at 1/2000 dilution
Predicted band size: 55 kDa -
Lanes 1 & 3 : Anti-ERG antibody [EPR3864] (ab92513) at 1/250 dilution (unpurified)
Lanes 2 & 4 : Anti-ERG antibody [EPR3864] (ab92513) at 1/1000 dilution (unpurified)
Lane 1 : Jurkat (human T cell leukemia cell line from peripheral blood) Whole Cell Lysate
Lanes 2-4 : Jurkat Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 55 kDa
Observed band size: 55 kDa
Exposure time: 12 minutes -
Immunohistochemical analysis of paraffin embedded Human Prostatic adenocarcinoma stage 3 tissue using unpurified ab92513 showing +ve staining.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (201)
ab92513 has been referenced in 201 publications.
- Zhu G et al. A novel peptide inhibitor of Dll4-Notch1 signalling and its pro-angiogenic functions. Br J Pharmacol 179:1716-1731 (2022). PubMed: 34796471
- Hattori Y et al. Embryonic Pericytes Promote Microglial Homeostasis and Their Effects on Neural Progenitors in the Developing Cerebral Cortex. J Neurosci 42:362-376 (2022). PubMed: 34819341
- Cheaito K et al. Establishment and characterization of prostate organoids from treatment-naïve patients with prostate cancer. Oncol Lett 23:6 (2022). PubMed: 34820005
- Fang Y et al. Epithelial Wntless regulates postnatal alveologenesis. Development 149:N/A (2022). PubMed: 34931663
- Facchinello N et al. Oxidative pentose phosphate pathway controls vascular mural cell coverage by regulating extracellular matrix composition. Nat Metab 4:123-140 (2022). PubMed: 35102339