ENO2 Antibody

Enolase-2, Mouse Anti Human
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In Stock
Cat. No.
BT16783
Synonyms
Gamma-enolase, EC 4.2.1.11, 2-phospho-D-glycerate hydro-lyase, Neural enolase, Neuron-specific enolase, NSE, Enolase 2, ENO2.
Appearance
Sterile filtered colorless solution.
Usage
THE BioTek's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
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Description

Introduction to ENO2 Antibody

ENO2 Antibody, also known as anti-neuron-specific enolase (NSE) antibody, is a research-grade immunological reagent designed to detect the ENO2 protein (enolase 2), a glycolytic enzyme with specialized roles in neuronal and cancer cell metabolism. ENO2 is a homodimeric enzyme (47 kDa) primarily expressed in neurons, neuroendocrine cells, and various cancers, serving as a diagnostic and prognostic biomarker .

Basic Characteristics of ENO2 Antibody

ParameterDetailsSources
Host/IsotypeMouse (IgG1), Rabbit (IgG), or Mouse (IgG2b Kappa)
ReactivityHuman, Mouse, Rat (cross-reactivity varies by vendor)
ApplicationsWestern Blot (WB), Immunohistochemistry (IHC), Flow Cytometry (FC), ELISA
ImmunogenRecombinant ENO2 fragments (e.g., AA 251–433) or synthetic peptides
PurificationAffinity chromatography (antigen-specific or protein A/G)
Storage2–8°C (short-term) or -20°C (long-term) with stabilizers (e.g., NaN₃, BSA)

Key Notes:

  • Specificity: Some antibodies (e.g., Proteintech’s 55235-1-AP) show no cross-reactivity with ENO1 or ENO3 .

  • Dilution Guidelines:

    • WB: 1:500–1:4000

    • IHC: 1:20–1:2000

    • FC: 1:200–1:400

Western Blotting

  • Protocol:

    1. Block membranes with 5% BSA or milk.

    2. Incubate with primary antibody (1:500–1:4000) at 4°C overnight.

    3. Detect with HRP-conjugated secondary antibodies .

  • Observed Bands: ~47 kDa .

Immunohistochemistry

  • Antigen Retrieval:

    • Citrate buffer (pH 6.0) or TE buffer (pH 9.0) recommended .

  • Staining Patterns:

    • Cytoplasmic: ENO2 localization in neurons and tumor cells .

    • Membrane-associated: Observed in specific cancer subtypes (e.g., glioblastoma) .

Flow Cytometry

  • Usage: Detects ENO2 expression in neuroendocrine tumors or immune cell subpopulations .

Prognostic Significance in Cancers

Cancer TypeENO2 ExpressionClinical CorrelationSources
DLBCL (Lymphoma)HighPoor OS, increased M2 macrophage infiltration
HNSCC (Head/Neck)HighGlycolysis dependency, PKM2 nuclear translocation
ccRCC (Kidney)HighEMT promotion, immune checkpoint therapy resistance
PDAC (Pancreas)HighMetastasis, poor survival, IGF-1-driven deacetylation

Mechanistic Roles in Tumorigenesis

  • Glycolysis Regulation:

    • HNSCC: ENO2 stabilizes PKM2, enhancing glycolytic flux and CCND1-driven proliferation .

    • CRC: ENO2 knockdown reduces migration, invasion, and EMT via YAP1 inactivation .

  • Immune Microenvironment:

    • ccRCC: High ENO2 correlates with TME immunosuppression (e.g., M2 macrophages) .

Comparative Analysis of ENO2 Antibodies

SupplierHostIsotypeReactivityApplicationsKey Features
AbceptaMouseIgG1Human, MouseWB, IHC, FCMonoclonal, recombinant immunogen
ProteintechRabbitIgGHuman, MouseWB, IHC, ELISANo cross-reactivity with ENO1/3
Bio-TechneMouseIgG2bHuman, RatIHC, Protein ArrayValidated by HuProt™ Array
EncorbioRabbitPolyHuman, RatWB, IHC, IF/ICCFull-length recombinant immunogen

Challenges and Considerations

  • Cross-Reactivity: Ensure specificity for ENO2 vs. ENO1/3 .

  • Tissue-Specific Staining:

    • Positive Controls: Neuroendocrine tumors, brain tissue .

    • Negative Controls: Muscle tissue (low ENO2 expression) .

  • Preclinical Translation: ENO2 inhibitors (e.g., AP-III-a4) show efficacy in HNSCC models .

Future Directions

  • Biomarker Development: ENO2 as a predictor for immune checkpoint therapy response in ccRCC .

  • Therapeutic Targeting: Dual inhibition of ENO2 and glycolytic pathways in cancers .

Product Specs

Introduction
Neuron-specific enolase, also known as NSE, is a glycolytic isoenzyme found in central and peripheral neurons and neuroendocrine cells. Enolase-2 is released into the cerebrospinal fluid (CSF) following neural tissue injury. Neoplasms originating from neural or neuroendocrine tissue release Enolase-2 into the bloodstream. Enolase-2 serves as a valuable biomarker and has been detected in patients diagnosed with specific tumors, including neuroblastoma, small cell lung cancer, medullary thyroid cancer, carcinoid tumors, pancreatic endocrine tumors, and melanoma. ENO2 is one of three enolase isoenzymes found in mammals. The ENO2 isoenzyme is present in mature neurons and cells of neuronal origin. A transition from alpha enolase to gamma enolase takes place in neural tissue during the developmental stages of rats and primates.
Physical Appearance
Sterile, colorless solution.
Formulation
1 mg/ml in phosphate-buffered saline (PBS), pH 7.4, containing 10% glycerol and 0.02% sodium azide.
Storage Procedures
For storage up to 1 month, maintain at 4°C. For extended storage periods, store at -20°C. Avoid repeated freeze-thaw cycles.
Stability / Shelf Life
Stable for 12 months at -20°C and 1 month at 4°C.
Applications
This antibody has undergone testing by ELISA and Western blot analysis to ensure its specificity and reactivity. However, as applications may vary, it is recommended to titrate the reagent for each specific experiment to achieve optimal results. The suggested dilution range for Western blot analysis is 1:500 to 1:5000, with an initial starting dilution of 1:1000.
Synonyms
Gamma-enolase, EC 4.2.1.11, 2-phospho-D-glycerate hydro-lyase, Neural enolase, Neuron-specific enolase, NSE, Enolase 2, ENO2.
Purification Method
ENO2 antibody was purified from mouse ascitic fluids by protein-A affinity chromatography.
Type
Mouse Anti Human Monoclonal.
Clone

PAT17D10A

Immunogen
Anti-human ENO2 mAb, clone PAT17D10A, is derived from hybridization of mouse F0 myeloma cells with spleen cells from BALB/c mice immunized with a recombinant human ENO2 protein 1-434 amino acids  purified from E. coli.
Ig Subclass
Mouse IgG2b heavy chain and Kappa light chain.

Product Science Overview

Structure and Function

Enolase-2 is one of the three isoforms of enolase, the others being enolase-1 (α-enolase) and enolase-3 (β-enolase). The enzyme exists as a homodimer or heterodimer, with the γγ homodimer being specific to neurons . The enzymatic site spans most of the length of the molecule, and it is highly conserved across species .

Clinical Significance

Elevated levels of Enolase-2 are often observed in various neuroendocrine tumors, including neuroblastoma and small-cell lung cancer . It is also used as a biomarker for neuronal damage and is measured in conditions such as stroke and traumatic brain injury .

Mouse Anti Human Enolase-2 Antibody

The Mouse Anti Human Enolase-2 Antibody is a monoclonal antibody that specifically targets the human Enolase-2 protein. This antibody is produced by immunizing mice with a synthetic peptide corresponding to the sequence of human Enolase-2 . It is widely used in research for detecting Enolase-2 in various applications, including Western blotting, immunohistochemistry, and flow cytometry .

Applications
  1. Western Blotting: Used to detect endogenous levels of Enolase-2 protein in cell extracts.
  2. Immunohistochemistry: Helps in visualizing the localization of Enolase-2 in tissue sections.
  3. Flow Cytometry: Used for intracellular staining to measure Enolase-2 levels in cells .
Species Reactivity

The Mouse Anti Human Enolase-2 Antibody shows cross-reactivity with human, mouse, and rat Enolase-2 . This makes it a versatile tool for studying Enolase-2 in various model organisms.

Storage and Handling

The antibody is typically supplied in a buffer containing sodium HEPES, NaCl, BSA, and glycerol. It should be stored at -20°C and should not be aliquoted to maintain its stability .

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