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Purified anti-ENO1 Antibody

Pricing & Availability
Clone
A18166G (See other available formats)
Regulatory Status
RUO
Other Names
Enolase 1; Alpha-Enolase; 2-Phospho-D-Glycerate Hydro-Lyase; Plasminogen-Binding Protein; Phosphopyruvate Hydratase
Isotype
Mouse IgG2b, κ
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Product Citations
publications
A18166G_PURE_ENO1_Antibody_1_03312021
Whole cell extracts (15 µg total protein) from indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 0.05 µg/mL purified anti-ENO1 antibody (clone A18166G) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG antibody (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:50000 dilution (lower). Lane M: Molecular weight marker.
  • A18166G_PURE_ENO1_Antibody_1_03312021
    Whole cell extracts (15 µg total protein) from indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 0.05 µg/mL purified anti-ENO1 antibody (clone A18166G) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG antibody (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:50000 dilution (lower). Lane M: Molecular weight marker.
  • A18166G_PURE_ENO1_Antibody_2_03312021
    Whole cell extracts (15 µg protein) from HeLa cells transfected with non-targeting control siRNA (siCon) or siRNA targeting ENO1 (siENO1) were resolved on a 4-12% Bis-Tris gel, transferred to PVDF and probed with 1.0 µg /mL (1:500 dilution) purified anti-ENO1 antibody (clone A18166G), overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG antibody (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:50000 dilution (lower). Lane M: Molecular Weight marker.
  • A18166G_PURE_ENO1_Antibody_3_03312021
    HeLa cells were fixed with ice-cold methanol for 10 minutes and blocked with 5% FBS for 60 minutes. Cells were then intracellularly stained with 5.0 µg /mL purified mouse IgG2b, κ isotype control antibody (Cat. No. 400302) (panel A) or 1.0 µg/mL (1:500 dilution) purified anti-ENO1 antibody (clone A18166G) (panel B) overnight at 4°C, followed by incubation with Alexa Fluor® 594 goat anti-mouse IgG antibody (Cat. No. 405326) at a 1:200 dilution. Nuclei were counterstained with DAPI and the image was captured with a 60X objective.
  • A18166G_PURE_ENO1_Antibody_4_03312021
    Whole cell extracts (250 µg total protein) prepared from HeLa cells were immunoprecipitated overnight with 2.5 µg of purified mouse IgG2b, κ isotype control antibody (Cat. No. 400302) or purified anti-ENO1 antibody (clone A18166G). 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 rabbit control antibody against a separate epitope of ENO1. Lane M: Molecular weight marker.
Cat # Size Price Quantity Check Availability Save
945501 25 µg 118 CHF
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945502 100 µg 293 CHF
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Description

Enolase is highly conserved, multifunctional protein. In the cytoplasm, ENO1 acts as a metabolic enzyme and catalyzes 2-phosphoglycerate to phosphoenolpyruvate in the last step of the glycolytic cycle.

Despite the lack of signal sequence in the ENO1 gene, ENO1 is found to be expressed on the surface of several hematopoietic cells though the transport mechanism is still unknown. Several of these cells include monocytes, T cells and B cells, and endothelial cells. On the cell surface, ENO1 acts as a plasminogen receptor and concentrates plasmin proteolytic activity at the pericellular area and protects plasmin from α2-antiplasmin inhibitors.

In cancer cells, ENO1 promotes what is known as the Warburg effect, a modified cellular metabolism in which the tumor cells uptake excessive glucose and undergo glycolysis to produce lactate in both hypoxic conditions and normal oxygen levels. ENO1 also promotes tumor invasion through plasminogen activation and extracellular matrix degradation.

Enolase can also be found in neurons and peripheral neuroendocrine cells. This neuron-specific enolase exists as ENO2 homodimer or an ENO1 and ENO2 heterodimer. Raised serum levels of these neuron-specific enolase have been found in all stages of neuroblastoma and it has been proposed that the plasmin formation caused by ENO1 enhances neuritogenesis.

The ENO1 gene can also be alternatively spliced and produce a 37 kD protein named c-myc promoter-binding protein, or MBP-1. MBP-1 is localized to the nucleus and binds to the c-myc P2 promoter, preventing the transcription of proto-oncogenes.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Recombinant human ENO1 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
KO/KD-WB, ICC, IP - 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 0.05 - 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 2.5 µg/test. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

When tested for ICC using 4%-PFA fixed HeLa cells permeabilized with methanol or Triton X-100, high non-specific staining in the nucleus was observed. We therefore do not recommend these fix-perm methods for this clone.

RRID
AB_2892513 (BioLegend Cat. No. 945501)
AB_2892513 (BioLegend Cat. No. 945502)

Antigen Details

Structure
ENO1 is a 434 amino acid protein with a predicted molecular weight of 47 kD.
Distribution

Cytosolic/Ubiquitously expressed

Function
Glycolytic enzyme
Biology Area
Cell Biology
Antigen References
  1. Capello M, et al. 2011. FEBS J. 278:1064-74
  2. Capello M, et al. 2016. Oncotarget. 7:5598-5612
  3. Cappello P, et al. 2017. Front Biosci (Landmark Ed). 22:944-959
  4. Díaz-Ramos A, et al. 2012. J Biomed Biotechnol. 2012:156795
  5. Didiasova M, et al. 2019. Front Cell Dev Biol. 7:61
  6. Isgrò MA, et al. 2015. Adv Exp Med Biol. 867:125-43.
  7. Liberti MV and Locasale JW. 2016. Trends Biochem Sci. 41:211-218.
Gene ID
2023 View all products for this Gene ID
UniProt
View information about ENO1 on UniProt.org

Other Formats

View All ENO1 Reagents Request Custom Conjugation
Description Clone Applications
Purified anti-ENO1 A18166G WB,KO/KD-WB,ICC,IP
Go To Top Version: 1    Revision Date: 03.31.2021

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.

 

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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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