Recombinant Anti-ENO1 antibody [EPR19758] (ab227978)

Product name

Anti-ENO1 antibody [EPR19758]
See all ENO1 primary antibodies
Description

Rabbit monoclonal [EPR19758] to ENO1
Host species

Rabbit
Specificity

ab227978 does not cross-react with ENO2 or ENO3.
Tested applications

Suitable for: IHC-P, WB, ICC/IF, IP, 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.
Positive control
- WB: MCF7, RAW 264.7, Jurkat, HeLa, NIH/3T3 and C6 whole cell lysates; human skeletal muscle tissue lysate; mouse brain, heart and kidney tissue lysates; rat brain, heart, liver and spleen tissue lysates. IP: Jurkat whole cell lysate. IHC-P: Human pancreas and clear cell renal cell carcinoma tissue; mouse and rat pancreas tissue. ICC/IF: HeLa and NIH/3T3 cells.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19758
Isotype

IgG
Research areas

Alternative Versions
- Anti-ENO1 antibody [EPR19758] - BSA and Azide free (ab229378)
Compatible Secondaries
Immunohistochemistry reagents
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant human ENO1 protein (Active) (ab89248)
Related Products
- Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab227978 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
IHC-P		1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB		1/1000. Predicted molecular weight: 47 kDa.
ICC/IF		1/500.
IP		1/30.
Flow Cyt (Intra)		1/600.

Notes
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Predicted molecular weight: 47 kDa.
ICC/IF 1/500.
IP 1/30.
Flow Cyt (Intra) 1/600.

Function

Multifunctional enzyme that, as well as its role in glycolysis, plays a part in various processes such as growth control, hypoxia tolerance and allergic responses. May also function in the intravascular and pericellular fibrinolytic system due to its ability to serve as a receptor and activator of plasminogen on the cell surface of several cell-types such as leukocytes and neurons. Stimulates immunoglobulin production.
MBP1 binds to the myc promoter and acts as a transcriptional repressor. May be a tumor suppressor.
Tissue specificity

The alpha/alpha homodimer is expressed in embryo and in most adult tissues. The alpha/beta heterodimer and the beta/beta homodimer are found in striated muscle, and the alpha/gamma heterodimer and the gamma/gamma homodimer in neurons.
Pathway

Carbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 4/5.
Sequence similarities

Belongs to the enolase family.
Developmental stage

During ontogenesis, there is a transition from the alpha/alpha homodimer to the alpha/beta heterodimer in striated muscle cells, and to the alpha/gamma heterodimer in nerve cells.
Post-translational
modifications

ISGylated.
Cellular localization

Nucleus and Cytoplasm. Cell membrane. Cytoplasm > myofibril > sarcomere > M line. Can translocate to the plasma membrane in either the homodimeric (alpha/alpha) or heterodimeric (alpha/gamma) form. ENO1 is localized to the M line.
Target information above from: UniProt accession P06733 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 2023 Human
- Entrez Gene: 100045967 Mouse
- Entrez Gene: 100503183 Mouse
- Entrez Gene: 13806 Mouse
- Entrez Gene: 433182 Mouse
- Entrez Gene: 24333 Rat
- Omim: 172430 Human
- SwissProt: P06733 Human
- SwissProt: P17182 Mouse
- SwissProt: P04764 Rat
- Unigene: 517145 Human
- Unigene: 372357 Mouse
- Unigene: 372389 Mouse
- Unigene: 70666 Mouse
- Unigene: 4236 Rat
see all
Alternative names
- 2 phospho D glycerate hydro lyase antibody
- 2-phospho-D-glycerate hydro-lyase antibody
- Alpha enolase antibody
- Alpha enolase like 1 antibody
- Alpha-enolase antibody
- C myc promoter binding protein antibody
- C-myc promoter-binding protein antibody
- EC 4.2.1.11 antibody
- eno1 antibody
- ENO1L1 antibody
- ENOA_HUMAN antibody
- Enolase 1 (alpha) antibody
- Enolase 1 (alpha) like 1 antibody
- Enolase 1 antibody
- Enolase alpha antibody
- MBP 1 antibody
- MBP-1 antibody
- MBP1 antibody
- MBPB1 antibody
- MPB 1 antibody
- MPB-1 antibody
- MPB1 antibody
- MYC promoter binding protein 1 antibody
- NNE antibody
- Non neural enolase antibody
- Non-neural enolase antibody
- Phosphopyruvate hydratase antibody
- Plasminogen binding protein antibody
- Plasminogen-binding protein antibody
- PPH antibody
- Tau crystallin antibody
see all

Western blot - Anti-ENO1 antibody [EPR19758] (ab227978)

All lanes : Anti-ENO1 antibody [EPR19758] (ab227978) at 1/1000 dilution

Lane 1 : Mouse brain tissue lysate
Lane 2 : Mouse heart tissue lysate
Lane 3 : Mouse kidney tissue lysate
Lane 4 : Rat brain tissue lysate
Lane 5 : Rat heart tissue lysate
Lane 6 : Rat liver tissue lysate
Lane 7 : Rat spleen tissue lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Predicted band size: 47 kDa
Observed band size: 47 kDa

Exposure time: 3 minutes

Blocking and dilution buffer: 5% NFDM/TBST.
Immunocytochemistry/ Immunofluorescence - Anti-ENO1 antibody [EPR19758] (ab227978)

Immunofluorescent analysis of 4% paraformaldegyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling ENO1 with ab227978 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and weakly nuclear staining in HeLa cell line.

The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

-ve control: PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution .
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENO1 antibody [EPR19758] (ab227978)

Immunohistochemical analysis of paraffin-embedded human clear cell renal cell carcinoma tissue labeling ENO1 with ab227978 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous and nuclear staining in human clear cell kidney carcinoma (PMID: 26037892) is observed. Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Flow Cytometry (Intracellular) - Anti-ENO1 antibody [EPR19758] (ab227978)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling ENO1 with ab227978 at 1/600 dilution (red) compared witha Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue).

Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-ENO1 antibody [EPR19758] (ab227978)

ENO1 was immunoprecipitated from 0.35 mg of Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate with ab227978 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab227978 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: Jurkat whole cell lysate 10 μg (Input).

Lane 2: ab227978 IP in Jurkat whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab227978 in Jurkat whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 3 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENO1 antibody [EPR19758] (ab227978)

Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling ENO1 with ab227978 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic and nuclear staining in medium-small ducts of human pancreas (PMID: 19425054) is observed. Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENO1 antibody [EPR19758] (ab227978)

Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue labeling ENO1 with ab227978 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic and nuclear staining in medium-small ducts of mouse pancreas is observed. Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENO1 antibody [EPR19758] (ab227978)

Immunohistochemical analysis of paraffin-embedded rat pancreas tissue labeling ENO1 with ab227978 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic and nuclear staining in medium-small ducts of rat pancreas is observed. Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Western blot - Anti-ENO1 antibody [EPR19758] (ab227978)

All lanes : Anti-ENO1 antibody [EPR19758] (ab227978) at 1/1000 dilution

Lane 1 : Human skeletal muscle tissue lysate at 20 µg
Lane 2 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate at 20 µg
Lane 3 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Lane 5 : C6 (rat glial tumor glial cell line) whole cell lysate at 10 µg
Lane 6 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg
Lane 7 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate at 10 µg

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Predicted band size: 47 kDa
Observed band size: 47 kDa

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure times.

Lane 1: 15 seconds.

Lanes 2-7: 1 second.
Immunocytochemistry/ Immunofluorescence - Anti-ENO1 antibody [EPR19758] (ab227978)

Immunofluorescent analysis of 4% paraformaldegyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryo fibroblast cell line) cells labeling ENO1 with ab227978 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and nuclear staining in NIH/3T3 cell line.

The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

-ve control: PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluo^r® 488) (ab150077) secondary antibody at 1/1000 dilution .
Anti-ENO1 antibody [EPR19758] (ab227978)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab227978? Please let us know so that we can cite the reference in this datasheet.

ab227978 has been referenced in 7 publications.

Xu W et al. m6A RNA methylation-mediated NDUFA4 promotes cell proliferation and metabolism in gastric cancer. Cell Death Dis 13:715 (2022). WB ; Human . PubMed: 35977935
Shi M et al. APCCDC20-mediated degradation of PHD3 stabilizes HIF-1a and promotes tumorigenesis in hepatocellular carcinoma. Cancer Lett 496:144-155 (2021). PubMed: 33039559
Swovick K et al. Interspecies Differences in Proteome Turnover Kinetics Are Correlated With Life Spans and Energetic Demands. Mol Cell Proteomics 20:100041 (2021). PubMed: 33639418
Yukimoto R et al. Specific activation of glycolytic enzyme enolase 2 in BRAF V600E-mutated colorectal cancer. Cancer Sci 112:2884-2894 (2021). PubMed: 33934428
Alam H et al. KMT2D Deficiency Impairs Super-Enhancers to Confer a Glycolytic Vulnerability in Lung Cancer. Cancer Cell 37:599-617.e7 (2020). PubMed: 32243837
Liu J et al. The circ-AMOTL1/ENO1 Axis Implicated in the Tumorigenesis of OLP-Associated Oral Squamous Cell Carcinoma. Cancer Manag Res 12:7219-7230 (2020). PubMed: 32884340
Yang T et al. Enolase 1 regulates stem cell-like properties in gastric cancer cells by stimulating glycolysis. Cell Death Dis 11:870 (2020). PubMed: 33067426

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Immunohistochemistry reagents

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Pathway

Sequence similarities

Developmental stage

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (7)

Customer reviews and Q&As

Post-translational
modifications