Enolase 1 Antibody (253) - BSA Free

Novus Biologicals | Catalog # NBP2-25147

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat, Porcine, Bovine, Equine

Cited:

Human

Applications

Validated:

Immunohistochemistry, Western Blot, Immunoassay, Immunocytochemistry/ Immunofluorescence, Simple Western

Cited:

Western Blot, Immunoassay

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 253

Format

BSA Free
View all Enolase 1 Primary Antibodies »

Product Specifications

Immunogen

The N-terminal 12 amino acids of bovine Enolase 1, which was synthesized on a 8-amine lysine core. [UniProt# Q9XSJ4]

Localization

Cytoplasm. Cell membrane. Cytoplasm > myofibril > sarcomere > M line.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Theoretical MW

47 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for Enolase 1 Antibody (253) - BSA Free

Western Blot: Enolase 1 Antibody (253) [NBP2-25147]

Western Blot: Enolase 1 Antibody (253) [NBP2-25147]

Western Blot: Enolase 1 Antibody (253) [NBP2-25147] - Analysis of different cell lysates using mouse mAb to alpha-enolase, NBP2-25147, dilution 1:10,000 in green: [1] protein standard (red), [2] NIH-3T3 l, [3] C6, [4] HEK293, [5] HeLa, and [6] SH-SY5Y cells. A strong single band at 47kDa corresponds to the alpha-enolase protein.
Immunocytochemistry/ Immunofluorescence: Enolase 1 Antibody (253) [NBP2-25147]

Immunocytochemistry/ Immunofluorescence: Enolase 1 Antibody (253) [NBP2-25147]

Immunocytochemistry/Immunofluorescence: Enolase 1 Antibody (253) [NBP2-25147] - Analysis of HeLa cells stained with mouse mAb to alpha-enolase, NBP2-25147, dilution 1:500 in green and costained with chicken pAb to HSP60, dilution 1:5,000, in red. The blue is DAPI staining of nuclear DNA. NBP2-25147 antibody reveals strong cytoplasmic staining.
Immunocytochemistry/ Immunofluorescence: Enolase 1 Antibody (253) [NBP2-25147]

Immunocytochemistry/ Immunofluorescence: Enolase 1 Antibody (253) [NBP2-25147]

Immunocytochemistry/Immunofluorescence: Enolase 1 Antibody (253) [NBP2-25147] - Rat 3T3 cells stained with NBP2-25147 (green) and counterstained with chicken polyclonal antibody to Vimentin NB300-223 (red) and DNA (blue). NBP2-25147 reveals strong cytoplasmic staining, while the Vimentin antibody reveals cytoplasmic intermediate filaments.
Simple Western: Enolase 1 Antibody (253) [NBP2-25147]

Simple Western: Enolase 1 Antibody (253) [NBP2-25147]

Simple Western: Enolase 1 Antibody (253) [NBP2-25147] - Simple Western lane view shows a specific band for Enolase 1 in 0.5 mg/mL of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Applications for Enolase 1 Antibody (253) - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:2000 - 1:5000

Immunohistochemistry

1:2000 - 1:5000

Simple Western

1:500

Western Blot

1:5000 - 1:10000
Application Notes
This Enolase 1 (253) antibody is useful for Immunocytochemistry/Immunofluorescence and Western blot, where a band can be seen at approx. 47 kDa. Use in Immmunoassay reported in scientific literature (PMID: 31035430).

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in HeLa lysate 0.5 mg/mL, separated by Size, antibody dilution of 1:500, apparent MW was 51 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

50% PBS, 50% glycerol

Format

BSA Free

Preservative

5mM Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Enolase 1

Enolases are enzymes which catalyze the conversion of 2-phosphoglycerate to phosphoenolpyruvate in the glycolytic pathway, and also the reverse reaction in gluconeogenesis. The Enzyme Commission classification number for this class of enzyme is EC4.2.1.11, and they can also be referred to as 2-phospho-D-glycerate hydrolases. There are three enolase proteins each coded for by a distinct gene. They are known as Enolase 1, 2 and 3 or alternately as alpha, beta and gamma-Enolase. Confusingly, although Enolase 1 is alpha-Enolase, Enolase 2 corresponds to gamma-Enolase while Enolase 3 is also known as beta-Enolase. All three enzymes exist as dimers in vivo and the three molecules are very similar in primary amino acids sequence. This similarity allows not only homodimers but also most possible heterodimers to form. The expression pattern of the three enzymes differs in a tissue specific manner, so that antibodies to them are useful cell type specific markers, especially as all three molecules are very abundant. Enolase 1 or alpha is also known as non-neuronal Enolase (NNE) and is expressed in most kinds of tissue, but is absent from neurons. Abnormal expression of NNE is associated with tumor progression in some breast and head and neck cancer. Enolase 2 or gamma is also known as neuron specific enolase (NSE). A switch from NNE to NSE occurs in the development of neurons. Enolase 3 or beta is expressed primarily in muscle cells.

Alternate Names

2-phospho-D-glycerate hydro-lyase, Alpha enolase, C-myc promoter-binding protein, c-myc promoter-binding protein-1, EC 4.2.1, EC 4.2.1.11, ENO1L1, Enolase 1, enolase 1, (alpha), MBP-1, MBPB1, MPB-1, MPB1c-myc promoter-binding protein 1, MYC promoter-binding protein 1, NNE, Non-neural enolase, Phosphopyruvate hydratase, Plasminogen-binding protein, PPHalpha enolase like 1, tau-crystallin

Entrez Gene IDs

2023 (Human)

Gene Symbol

ENO1

OMIM

172430 (Human)

UniProt

P06733 (Human)

Additional Enolase 1 Products

Product Documents for Enolase 1 Antibody (253) - BSA Free

Certificate of Analysis

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Product Specific Notices for Enolase 1 Antibody (253) - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for Enolase 1 Antibody (253) - BSA Free

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Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for Enolase 1 Antibody (253) - BSA Free

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  • Q: Enolase came up as a candidate in mass spec. There are at least three isoforms which showed up once or twice or three times (as in three mass spec repeats). To validate the mass data, we are looking into quantitate each isoform. Which antibodies are specific for each of the three?

    A: We checked all antibodies to Enolase 1, Enolase 2 and Enolase 3, but unfortunately no cross reactivity testing was performed for any of these antibodies. Considering how similar these proteins are, it is reasonable to expect some of them will react with other enolases. Base on immunogen sequence, we found some that were raised to the most diverse regions between these proteins, so it is possible that these will be the most  specific to the protein they were raised to.

    For Enolase 1, your best bet is NBP2-25147  as it was raised to N terminal peptide of bovine Eno1: MSILKVHAREIF

    For Enolase 2, I found 3 candidate antibodies:
    NBP2-53158  and NBP2-50532  were raised to aa416-433 (ELGDEARFAGHNFRNPSVL) - it says these antibodies are only reactive with human but they will also work for mouse since the immunogen is 100% match
    NBP2-50533, which was raised to aa271-285 (GDQLGALYQDFVRD) - which is also likely to react with mouse, the immunogen is 93% match with mouse (only last aminoacid is different)

    For Enolase 3, your best bet would be H00002027-M01 or H00002027-A01. both were raised to aa228-277 (KTAIQAAGYPDKVVIGMDVAASEFYRNGKYDLDFKSPDDPARHITGEKLG) - 98% match to mouse

    The immunogens for all these antibodies fall into most variable regions, however, since we have not performed crossreactivity testing, we can not guarantee that they will not crossreact to some extent.

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