Anti-Enolase 1 (ENO1, 2-phospho-D-glycerate Hydrolyase, alpha Enolase, C-myc Promoter-binding Protei

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 refered 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, as can be seen from a sequence alignment downloadable from here. 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 (1, 2). Enolase 2 or gamma is also known as neuron specific enolase (NSE), and we also market antibodies directed against this protein, specifically 031410. A switch from NNE to NSE occurs in the development of neurons (3). Enolase 3 or beta is expressed primarily in muscle cells. Monoclonal antibody 031356 was raised against the N-terminal 12 amino acids of bovine enolase 1 which was synthesized on a 8-amine lysine core using the multiple antigen presentation method (5). This produces a dendrimer presenting 8 peptides to the immune system. The characterization of the antibody has been published in peer reviewed form and in this publication is there referred to as Enol2-53 (4). The antibody was tested for binding to expressed bovine enolase 1, 2 and 3 and shown to be specific for only enolase 1. The HGNC name for this protein is ENO1. Applications: Suitable for use in Immunofluorescence and Western Blot. The NNE protein runs at about ~47kD on SDS-PAGE gels. Other applications not tested. Recommended Dilution: Immunofluorescence: 1:500-1:1000, Western Blot: 1:5000, Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.