Product Name:
ENO2-pY25
Product Number:
ab-pn512
Target Full Name: Gamma-enolase
Target Alias: 2-phospho-D-glycerate hydro-lyase; 2-phospho-D-glycerate hydrolyase; ENO2; ENOG; Enolase 2; enolase 2 (gamma, neuronal); Gamma-enolase; Neural enolase; neuron specific gamma enolase; Neuron-specific enolase; neurone-specific enolase; NSE
Product Type Specific: ENO2 phosphosite-specific antibody
Antibody Code: PN512
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: Y25
Protein UniProt: P09104
Protein SigNET: ENO2
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Human ENO2 sequence peptide Cat. No.: PE-04ACT95
Antibody Immunogen Sequence: VDL(pY)TAK(bA)C
Antibody Immunogen Description: Corresponds to amino acid residues V22 to K28; in the Enolase_N domain. The effect of Y25 phosphorylation is unclear. This is one of the major in vivo phosphorylation sites in ENO2 (≥100 reports from high throughput mass spectrometry studies recorded in PhosphoSitePlus).
Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.This antibody was also subject to negative purification over phosphotyrosine-agarose.
Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.
Antibody Concentration: 1 mg/ml
Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol
Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.
Product Use: Western blotting | Antibody microarray
Antibody Dilution Recommended: 2 µg/ml for immunoblotting
Antibody Potency: Medium immunoreactivity with immunogen peptide on dot blots.
Antibody Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Rhesus Macaque | Mouse | Platypus | Chicken | Frog.
Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 45-55 kDa.
Antibody Specificity: High-very high
Antibody Cross Reactivity: No significant cross-reactivities detected in A431 and A549 cells, although background is a little high with EGF treatment of A549 cells
Related Product 1: ENO2 (22-28) pY25 antibody blocking peptide (Cat. No.: PE-04ACT95)
Related Product 2: ENO1 Y44 phosphorylation antibody (Cat. No.: AB-PN511)
Scientific Background: ENO2 (Enolase 2 or Neuron-Specific Enolase, NSE) is a glycolytic enzyme (434-amino acids) that catalyzes the conversion of 2-phosphoglycerate to phosphoenolpyruvate. It is crucial for neuronal energy metabolism and survival, particularly in neurons and neuroendocrine cells. ENO2 acts as a neurotrophic factor that promotes neuronal cell survival. It plays a part in various processes such as growth control, hypoxia tolerance and allergic responses. As a receptor and activator of plasminogen on the cell surface of several cell-types such as leukocytes and neurons, it also functions in the intravascular and pericellular fibrinolytic system. It functions as a homodimer of two identical alpha subunits, but can exist as heterodimers in some tissues. It is located in cytoplasm (glycolysis), the cell surface (plasminogen receptor), the nucleus (gene regulation), and centrosomes. It is released into CSF or blood upon neural damage, with significant elevation during brain trauma, cerebral accidents, and neuroendocrine tumours. It is often overexpressed in various cancers (e.g., neuroblastoma, melanoma, small cell lung cancer, glioblastoma, thyroid, pancreatic) and associated with poor prognosis. This description may include information annotated by UniProt and/or Google AI.