Eosinophil Derived Neurotoxin (EDN) ELISA Assay Kit

$990.00

The Eagle Biosciences Eosinophil Derived Neurotoxin (EDN) ELISA Assay Kit is intended for the quantitative determination of eosinophil derived neurotoxin (EDN) in stool, plasma, serum and urine. The EDN ELISA Assay Kit is for Research Use Only and is not for use in diagnostic procedures.

SKU: EDN39-K01 Categories: , ,

Eosinophil Derived Neurotoxin (EDN) ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 0.164 ng/mL
Incubation Time: 2.5 hours
Sample Type: Stool, Urine, Serum, Plasma
Sample Size: 100 µl
Controls Included

Additional Information

Assay Background

After activation eosinophil granulocytes segregate the cationic glycoprotein EDN (eosinophil derived neurotoxin). This 18-21 kDa single stranded glycosylated protein is also known as EPX (eosinophil protein X). Together with ECP (eosinophil cationic protein), EDN belongs to the ribonuclease superfamily1-3. EDN, however, has a 100-fold increased ribonuclease activity. EDN is also neurotoxic and not cytotoxic4,5. The activation of eosinophil granulocytes is important during the inflammatory processes in allergic reactions. Thus EDN is a marker for eosinophil activation and degranulation.

The measurement of EDN in stool allows the detection of clinical or subclinical chronic inflammation in the gut. Some research has suggested that the measurement of EDN gives information on the activity of disease and the prediction of a relapse. The Eosinophil Derived Neurotoxin (EDN) ELISA Assay Kit allows an easy, rapid and precise quantitative determination of the eosinophil derived neurotoxin in biological samples. The kit includes all reagents ready to use for preparation of the samples.

Assay Principle

The Eosinophil Derived Neurotoxin (EDN) ELISA Assay Kit determines human EDN according to the “sandwich” ELISA principle. EDN in sample, standard and controls binds to monoclonal antibodies, which are coated to the microtiter plate. After a washing step, a peroxidase labeled polyclonal antibody is added. A second washing step is followed by the addition of the substrate which is converted to a colored product by the peroxidase. The reaction is terminated by the addition of an acidic stop solution. The optical densities are read at 450 nm in a microtiter plate reader. The EDN concentration can be calculated from the standard curve.

1. Sample Incubation

  • Pipette 100µl STD (undiluted), CTRL (1:5 diluted with ASSYBUF) and samples in duplicate to the microtiter plate. For standard 0 use the corresponding dilution buffer (See Section 12) of the samples
  • The strips are covered and incubated by shaking for 60 min at room temperature (18-26 °C).
  • The reaction in each well starts immediately. Pipetting should be performed as quickly as possible. When processing many samples at once the samples should be pipetted to a separate microtiter plate (150 µl) and transferred simultaneously using a multichannel pipette.

2. Washing step

  • Discard the contents of the microwells and wash 5x with 250 µl diluted WASHBUF per well. Remove residual buffer by tapping the plate on absorbent paper after the last washing step.

3. Incubation conjugate

  • Pipette 100 µl CONJ in each microwell.
  • The strips are covered and incubated by shaking at room temperature (18-26 °C) for 60 min.

4. Washing step

  • Discard the contents of the microwells and wash 5x with 250 µl diluted WASHBUF per well. Remove residual buffer by tapping the plate on absorbent paper after the last washing step.

5. Incubation substrate

  • Pipette 100 µl SUB in each microwell.
  • Incubate for 5-10 min at room temperature (18-26 °C) in the dark.

6. Stopping reaction

  • Pipette 100 µl STOP in each microwell, mix well.

7. Reading

  • Read the absorbance at 450 nm. If the microtiter plate reader allows to use a reference wavelength use 620 or 690 nm as reference wavelength.
  • Reading should be done within 30 min after stopping reaction.
  • In case that the highest standard exceeds the range of the reader the reading should be done at 405 nm against 620 nm (690 nm).

Typical Standard Curve

Eosinophil Derived Neurotoxin (EDN) ELISA Assay Kit Standard Curve

Manual

Product Manual