NSE Monoclonal antibody

• Featured Product
• KD/KO Validated

NSE Monoclonal Antibody for WB, IHC, IF-P, FC (Intra), ELISA

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat, pig

Applications

WB, IHC, IF-P, FC (Intra), ELISA

Conjugate

Unconjugated

Publications

20

CloneNo.

6F8G3

Reviews (1)

Cat no : 66150-1-Ig

Synonyms

ENO2, Gamma enolase, Enolase 2, EC:4.2.1.11, 6F8G3

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Validation Data Gallery

• 

  WB analysis using 66150-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66150-1-Ig (NSE antibody) at dilution of 1:50000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated GAPDH Monoclonal antibody (HRP-60004) as loading control.

• 

  WB analysis of U-251 using 66150-1-Ig

  WB result of NSE antibody (66150-1-Ig; 1:4000; incubated at room temperature for 1.5 hours) with sh-Control and sh-NSE transfected U-251 cells.

• 

  WB analysis of Neuro-2a using 66150-1-Ig

  Neuro-2a cells were subjected to SDS PAGE followed by western blot with 66150-1-Ig (NSE antibody at dilution of 1:2000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 66150-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66150-1-Ig (NSE antibody) at dilution of 1:50000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of U-251 using 66150-1-Ig

  U-251 cells were subjected to SDS PAGE followed by western blot with 66150-1-Ig (NSE antibody at dilution of 1:4000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of fetal human brain using 66150-1-Ig

  fetal human brain tissue were subjected to SDS PAGE followed by western blot with 66150-1-Ig (NSE antibody at dilution of 1:4000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of rat brain using 66150-1-Ig

  rat brain tissue were subjected to SDS PAGE followed by western blot with 66150-1-Ig (NSE antibody at dilution of 1:4000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of pig brain using 66150-1-Ig

  pig brain tissue were subjected to SDS PAGE followed by western blot with 66150-1-Ig (NSE antibody at dilution of 1:4000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of SH-SY5Y using 66150-1-Ig

  SH-SY5Y cells were subjected to SDS PAGE followed by western blot with 66150-1-Ig (NSE antibody at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of U-251 using 66150-1-Ig

  U-251 cells were subjected to SDS PAGE followed by western blot with 66150-1-Ig (NSE antibody at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of C6 using 66150-1-Ig

  C6 cells were subjected to SDS PAGE followed by western blot with 66150-1-Ig (NSE antibody at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  IHC staining of mouse brain using 66150-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 66150-1-Ig (NSE antibody) at dilution of 1:5000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse brain using 66150-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 66150-1-Ig (NSE antibody) at dilution of 1:5000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human brain using 66150-1-Ig

  Immunohistochemical analysis of paraffin-embedded human brain tissue slide using 66150-1-Ig (NSE antibody at dilution of 1:200 (under 10x lens).

• 

  IHC staining of human brain using 66150-1-Ig

  Immunohistochemical analysis of paraffin-embedded human brain tissue slide using 66150-1-Ig (NSE antibody at dilution of 1:200 (under 40x lens).

• 

  IHC staining of rat cerebellum using 66150-1-Ig

  Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue slide using 66150-1-Ig (NSE antibody) at dilution of 1:5000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat cerebellum using 66150-1-Ig

  Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue slide using 66150-1-Ig (NSE antibody) at dilution of 1:5000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat brain using 66150-1-Ig

  Immunohistochemical analysis of paraffin-embedded rat brain tissue slide using 66150-1-Ig (NSE antibody) at dilution of 1:5000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat brain using 66150-1-Ig

  Immunohistochemical analysis of paraffin-embedded rat brain tissue slide using 66150-1-Ig (NSE antibody) at dilution of 1:5000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of mouse brain using 66150-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed mouse brain tissue using NSE antibody (66150-1-Ig, Clone: 6F8G3 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

• 

  IF Staining of mouse brain using 66150-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed mouse brain tissue using NSE antibody (66150-1-Ig, Clone: 6F8G3 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

• 

  FC experiment of SH-SY5Y using 66150-1-Ig

  1X10^6 SH-SY5Y cells were intracellularly stained with 0.2 ug Anti-Human NSE (66150-1-Ig, Clone:6F8G3) and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.2 ug Control Antibody. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: PC-12 cells, C6 cells, fetal human brain tissue, HEK-293 cells, Neuro-2a cells, pig brain tissue, rat brain tissue, SH-SY5Y cells, U-251 cells, pig cerebellum tissue, rat cerebellum tissue, mouse brain tissue, mouse cerebellum tissue
• Positive IHC detected in: mouse brain tissue, human brain tissue, rat cerebellum tissue, rat brain tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF-P detected in: mouse brain tissue
• Positive FC (Intra) detected in: SH-SY5Y cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:2500-1:10000
• Immunofluorescence (IF)-P: IF-P : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.20 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 1 publications below
• WB: See 11 publications below
• IHC: See 4 publications below
• IF: See 7 publications below

Product Information

66150-1-Ig targets NSE in WB, IHC, IF-P, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat, pig samples.

• Tested Reactivity: human, mouse, rat, pig
• Cited Reactivity: human, mouse, rat
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: NSE fusion protein Ag19106
• Full Name: enolase 2 (gamma, neuronal)
• Calculated Molecular Weight: 47 kDa
• Observed Molecular Weight: 47 kDa
• GenBank Accession Number: BC002745
• Gene Symbol: NSE
• Gene ID (NCBI): 2026
• RRID: AB_2881546
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

NSE, also named as ENO2, belongs to the enolase family. Enolases are cytoplasmic glycolytic enzymes that may be involved in differentiation. The enolase has three isoenzymes, alpha, beta and gamma. The alpha form is expressed in most tissues, whereas the beta form is expressed in muscle tissue. The gamma enolase (ENO2), a homodimer, is primarily localized in neurons and neuroendocrine cells and is a cancer diagnostic marker for brain tumors (PMID:7520111). ENO2 plays a role in the glycolysis-related energy pathway and might be involved in higher metabolic activity during the day than at night, at least in part.

Publications

Species	Application	Title
human	WB	Adv Sci (Weinh); Identification of PRDX5 as A Target for The Treatment of Castration-Resistant Prostate Cancer; Authors - Rong Wang
human	IF	Oncogenesis; Neuron-specific enolase promotes stem cell-like characteristics of small-cell lung cancer by downregulating NBL1 and activating the BMP2/Smad/ID1 pathway.; Authors - Lin Lu
rat	IHC	J Neurochem; Extracellular ATP and cAMP signaling promote Piezo2-dependent mechanical allodynia after trigeminal nerve compression injury.; Authors - Zhaoke Luo
rat	IF	CNS Neurosci Ther; Sema7A, a brain immune regulator, regulates seizure activity in PTZ-kindled epileptic rats.; Authors - Jing Deng
rat	IHC	Neuroreport; Effects of morphine preconditioning on TRPV1 sensitization and ERK1/2 phosphorylation induced by TGFβ1 in neurocytes.; Authors - Zhen-Xiao Ma
rat	IF	Biomed Res Int; In Vitro Neural Differentiation of Bone Marrow Mesenchymal Stem Cells Carrying the FTH1 Reporter Gene and Detection with MRI.; Authors - Tong Mu

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Morgan (Verified Customer) (10-26-2021)

Overall it worked well. It looked a little dirty with some "extra" bands, but I'm not exactly sure why. The trend is still clear though.

• Applications: Western Blot
• Primary Antibody Dilution: 1:500
• Cell Tissue Type: OV90 Ovarian Cancer Cells

NSE Antibody Western Blot validation (1:500 dilution) in OV90 Ovarian Cancer Cells (Cat no:66150-1-Ig)

Related Products

Flow Cytometry Perm Buffer (10X)

PF00011-C