Cancer Biomarker ELISA Assay

NSE ELISA Assay Kit

$585.00

The NSE ELISA Assay Kit (enzyme-linked immunoassay kit) is intended for the quantitative determination of human neuron specific enolase (NSE) levels in serum. The Eagle Biosciences Human Neuron Specific (NSE) ELISA Assay Kit is for research use only and not to be used in diagnostic procedures.

SKU: NSE31-K01 Categories: , ,

NSE ELISA Assay Kit

NSE ELISA Assay Kit Developed and Manufactured in the USA

Size: 1×96 wells
Sensitivity: 1.2 ng/mL
Dynamic Range: 5.0 – 178 ng/ml
Incubation Time: 1.0 hours
Sample Type: Serum
Sample Size: 10 µL
Alternative Names: Human Neuron Specific Enolase
For Research Use Only

Expected Normal Values:
One hundred seventy two normal adult sera were measured with this human Neuron Specific Enolase (NSE) ELISA. The normal range was found to be less than 15 ng/mL. It is highly recommend that each laboratory should establish its own normal cut off level.

Controls Included

Assay Principle

The NSE ELISA Assay Kit is designed, developed and produced for the quantitative measurement of human Neuron Specific Enolase in serum sample. The assay utilizes the two-site “sandwich” technique with two selected monoclonal antibodies that bind to different epitopes of the γ-subunit of the enzyme.

Assay standards, controls and samples are added directly to wells of microplate that is coated with a streptavidin. Subsequently, a mixture of a biotinylated NSE specific monoclonal antibody and a horseradish peroxidase (HRP) labeled NSE specific monoclonal antibody is added to each microtiter well. After the first incubation a “sandwich” immunocomplex of “streptavidin-biotin-monoclonal antibody – human NSE – monoclonal antibody-HRP” is formed. The unbound monoclonal antibodies are removed in the subsequent washing step. For the detection of this immunocomplex, the well is then incubated with a substrate solution in a timed reaction and then measured in a spectrophotometric microplate reader. The enzymatic activity of the immunocomplex bound to the NSE on the wall of the microtiter well is directly proportional to the amount of NSE in the sample. A standard curve is generated by plotting the absorbance versus the respective human NSE concentration for each standard on point-to-point, cubical scales or 4 parameter curve fit. The concentration of human NSE in test samples is determined directly from this standard curve.

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Additional Information

Assay Background

The glycolytic enzyme enolase (2-phospho-D-glycerate hydrolyase) exists as several dimeric isoenzymes (αα, αβ, ββ and γγ) composed of three distinct subunits: α, β, and γ. Three isoenzymes are found in human brain: αα, αβ, and γγ. The heterologous αγ-isoenzyme and the homologous γγ -enolase isoenzymes are known as neuron-specific enolase (NSE) as these isoenzymes initially were detected in neurons and neuroendocrine cells. By using monoclonal antibodies specific to the γ-subunit of the enzyme allow the test to detect both the αγ and the γγ forms.
The NSE levels are quite low in normal healthy people and in people with benign disease. Lung cancer is one of the most common cancer forms with incidences about 50-100 per 100,000 population. Approximately 20% of the lung cancer is small cell lung cancer. NSE has been shown to be a valuable tumor marker of neuroendocrine origin, particularly in small cell lung cancer and in neuroblastoma. Although NSE is similar to Chromogranin A in detecting small cell lung cancer and neuroblastoma, Chromogranin A seems better in detecting carcinoid.

Assay Procedure

  1. Prepare NSE Tracer Antibody and Capture Antibody working solution by 1:21 fold dilution of the Tracer Antibody with the biotinylated
  2. Capture Antibody. For each strip, it is required to mix 1 mL of the Capture Antibody with 50 µL of the Tracer Antibody in a clean test tube.
  3. Add 10 µL of standards, controls and samples into the designated microwell.
  4. Add 100 µL of above mixture of Tracer Antibody and Capture Antibody solution to each of the wells.
  5. Cover the plate with the plate sealer and incubate plate at room temperature, shaking at 170 rpm for 1 hour.
  6. Remove plate sealer. Aspirate the contents of each well. Wash each well 5 times by dispensing 350 µL of working wash solution into each well and then completely aspirating the contents. Alternatively, an automated microplate washer can be used.
  7. Add 100 µL of ELISA HRP Substrate into each of the wells.
  8. Cover the plate with one plate sealer and also with aluminum foil to avoid exposure to light.
  9. Incubate plate at room temperature for 10 minutes or less.
  10. Remove the aluminum foil and plate sealer. Add 100 µL of ELISA Stop Solution into each of the wells. Mix gently.
  11. Read the absorbance at 450 nm within 10 minutes in a microplate reader.

Typical Standard Curve

NSE ELISA Assay Kit

Documents

Product Documents

Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.

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