NSE, BioAssay(TM) ELISA Kit (Gamma-enolase, 2-phospho-D-glycerate Hydro-lyase, Neural Enolase, Neuron-specific Enolase, NSE, Enolase 2, ENO2, NSE)

Enolase (2-phosphogly-cerate hydrolyase or phosphopyruvate hydrates) is a glycolytic enzyme that catalyzes the dehydration and conversion of 2-phosphoglycerate to phosphoenolpyruvate. It comprises three distint subunits, a, b and y. The yy and ay dimeric forms of enolase, referred to as neuron-specific enolase(NSE), are localized mainly in neurons and neuroectodermal tissue. NSE has a high stability in biological fluids and can easily diffuse to the extracellular medium and cerebrospinal fluid(CSF) when neuronal membranes are injured. NSE is used clinically as a sensitive and useful marker of neuronal damage in several neurological disorders including stroke, hypoxic brain damage, status epilepticus, Creutzfeldt- Jakob disease, and herpetic encephalitis.\n\nIntended Use:\nHuman neuron specific enolase ELISA kit is to be used for the in vitro quantitative determination of human NSE in human serum, human plasma, cell lysate and buffered solution. The assay will recognize native NSE. This kit has been configured for research use only and is not to be used in diagnostic procedures.\n\nSensitivity:\nThe minimal detectable dose of human NSE was calculated to be 0.15ng/ml, by subtracting two standard deviations from the mean of 12 zero standard replicates (ELISA buffer, S0) and intersecting this value with the standard curve obtained in the same calculation.\n\nSpecificity:\nThe following substances were tested and found to have no cross-reactivity: human serum albumin, human non neuronal enolase, human alpha fetoprotein, human prostate specific antigen (PSA), human hemoglobin, human VDBP (vitamin D binding protein)\n\nTest Principle:\nThe design of this assay is based on a sandwich Enzyme-Linked Immunosorbent Assay (ELISA). The microtiter plate provided in this kit has been pre-coated with a monoclonal antibody specific to human NSE. Samples are pippetted into these wells. Nonbound NSE and other components of the sample should be removed by washing, then biotin-conjugated monoclonal antibody specific to NSE added. In order to quantitatively determine the amount of NSE present in the sample, Avidin conjugated to Horseradish Peroxidase (HRP) should be added to each microplate well. The final step, a TMB-substrate solution added to each well. Finally, a sulfuric acid solution is added and the resulting yellow colored product is measured at 450nm. Since the increases in absorbency is directly proportional to the amount of captured NSE.\n\nKit Components:\nMicroplate: 1x96 wells\nIncubation buffer: 1x30ml\nWashing buffer (10x): 1x100ml\nStandard protein: 1 glass vial lyophilized\nStandard/sample dilution buffer: 1x25ml\nSecond antibody: 1 glass vial lyophilized\nAV-HRP: 1x150ul\nSecondary antibody/AV-HRP dilution buffer: 1x25ml\nSubstrate (TMB): 1x20ml\nStop solution: 1x20ml\n\nStorage and Stability:\nStore components at 4 degrees C. Stable for at least 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

N2175-31

Size

96Tests

Applications

ELISA

References

1. Fletcher L. et al. (1976) Biochim. Biophys. Acta. 452(1), 245-252 2. Lima J.E. et al. (2004) J. Neurol. Sci. 217(1), 31-35 3. Suzuki Y. et al. (1999) Neurology 53(8), 1761-1764