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Purified anti-ERG Antibody

Pricing & Availability
Clone
W19256A (See other available formats)
Regulatory Status
RUO
Other Names
ETS-Related Gene, Transcriptional Regulator ERG, ETS Transcription Factor ERG, Erythroblast Transformation-Specific Transcription Factor ERG Variant 10, p55
Isotype
Rat IgG2a, κ
Ave. Rating
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Product Citations
publications
W19256A_PURE_ERG_Antibody_1_031621
Whole cell extracts (15 µg total protein) from MOLT-4 (high expressor) , HeLa (low expressor), and MCF7 (low expressor) and U937 (negative control) lysates were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 μg/mL (1:500 dilution) purified anti-ERG antibody (clone W19256A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-rat IgG antibody (Cat. No. 405405) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:25000 dilution (lower). Lane M: Molecular weight marker.
  • W19256A_PURE_ERG_Antibody_1_031621
    Whole cell extracts (15 µg total protein) from MOLT-4 (high expressor) , HeLa (low expressor), and MCF7 (low expressor) and U937 (negative control) lysates were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 μg/mL (1:500 dilution) purified anti-ERG antibody (clone W19256A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-rat IgG antibody (Cat. No. 405405) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:25000 dilution (lower). Lane M: Molecular weight marker.
  • W19256A_PURE_ERG_Antibody_2_031621
    Whole cell extracts (250 µg total protein) prepared from MOLT-4 cells were immunoprecipitated overnight with 2.0 µg of purified rat IgG2a, k isotype ctrl antibody (Cat. No. 400501) or purified anti-ERG antibody (clone W19256A). The resulting IP fractions and whole cell extract input (6%) were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane and probed with a control antibody against a separate epitope of ERG. Lane M: Molecular weight marker.
  • W19256A_PURE_ERG_Antibody_3_031621
    HeLa cells (negative control) (panel A) and MOLT-4 cells (positive control) (panel B) were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with Triton X-100 for 10 minutes, and blocked with 5% FBS for 60 minutes. Cells were then intracellularly stained with purified anti-ERG antibody (clone W19256A) overnight at 4°C followed by incubation with Alexa Fluor® 594 goat anti-rat IgG antibody (Cat. No. 405422) at 2.0 µg/mL. Nuclei were counterstained with DAPI and the image was captured with a 60X objective.
  • W19256A_PURE_ERG_Antibody_4_042121
    MOLT4 cells (filled histogram, positive control) or U937 cells (open histogram, negative control) were fixed and permeabilized using the True-Nuclear™ Transcription Factor Buffer set (Cat. No. 424401), and intracellularly stained with ERG (clone W19256A) followed by PE goat anti-rat IgG antibody (Cat. No. 405406).
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940201 25 µg $118
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940202 100 µg $293
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Description

ERG (ETS-related gene) is a member of the erythroblast transformation-specific (ETS) family of transcription factors. Expressed in the mesoderm and endothelium of embryos, ERG is vital to embryonic cell differentiation as well as regulation of endothelial homeostasis, a role it sustains beyond the embryonic stage. Through transcriptional regulation of VE-cadherin, ICAM-2, and Claudin-5, ERG maintains endothelial junctional integrity and stability and regulation of the endothelial cytoskeleton through its interactions with HDAC6 and RhoJ. ERG is vital to angiogenesis, bone development, platelet adhesion, urogenital tract development, vascular stability, and proper hematopoiesis regulation. It plays a large role in cellular response to vascular inflammation, endothelial cell barrier function, and inhibition of vascular inflammation. These functions are accomplished largely through ERG-mediation of the Wnt/β-catenin signaling pathway by means of transcriptional regulation of FZD4 and VE-cadherin by ERG and subsequent promotion of β-catenin stability. This in turn promotes endothelial cell proliferation. ERG fusion proteins resulting from chromosomal translocations cause ERG to behave as a proto-oncogene, contributing to prostate cancer, megakaryoblastic leukemia, and Ewing’s sarcoma.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Partial recombinant human ERG protein
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC, IP, ICFC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 0.125 - 1.0 µg/mL. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/mL is recommended. For immunoprecipitation, the suggested use of this reagent is 2.0 µg/test. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/mL is recommended. For intracellular flow cytometry using our True-Nuclear™ Transcription Factor Staining Protocol, the suggested use of this reagent is ≤ 0.25 µg per million cells in 100 µL volume. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

This clone was tested for ICC using HeLa (low expressorl) and MOLT-4 (high expressor) cells fixed with 4% paraformaldehyde and permeabilized with either methanol or Triton X-100. Both methods resulted in robust ERG staining.  

This clone does not recognize mouse ER.

When using this clone for ICFC, we recommend using the True-Nuclear™ Transcription Factor Buffer Set or the True Phos perm buffer. We do not recommend using the Intracellular Staining Permeabilization Wash Buffer for ICFC testing due to poor ERG staining.

True-Nuclear™ Transcription Factor Buffer Set gave a better signal to noise ratio at all tested concentrations when compared to the True Phos perm buffer.

RRID
AB_2892503 (BioLegend Cat. No. 940201)
AB_2892503 (BioLegend Cat. No. 940202)

Antigen Details

Structure
ERG is a 449 amino acid protein with an expected molecular weight of 54 kD.
Distribution

Ubiquitously expressed/ Nucleus 

Function
Transcription factor
Cell Type
Endothelial cells
Biology Area
Cell Biology, Ubiquitin/Protein Degradation
Antigen References

1. Adamo P, et al. 2016. J. Biol. Chem. 35:4013
2. Birdsey G, et al. 2015. Dev. Cell. 32:82
3. Shah A, et al. 2016. Vascul. Pharmacol. 86:3

 

Gene ID
2078 View all products for this Gene ID
UniProt
View information about ERG on UniProt.org
Go To Top Version: 2    Revision Date: 04/21/2021

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

BioLegend, the BioLegend logo, and all other trademarks are property of BioLegend, Inc. or their respective owners, and all rights are reserved.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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