Noradrenaline (Norepinephrine) Sensitive ELISA

Additional Information

Assay Background

The Noradrenaline (Norepinephrine) High Sensitive ELISA Assay kit is an Enzyme-linked immunosorbent assay used for the quantitative and very sensitive determination of noradrenaline in biological samples including serum, plasma, tissue, and cell culture samples.  The Noradrenaline (Norepinephrine) High Sensitive ELISA Assay kit is for research use only and should not be used in diagnostic procedures.  Catecholamine is the name of a group of aromatic amines (noradrenaline, adrenaline, dopamine, and their derivatives) which act as hormones and neurotransmitter, respectively. Adrenaline and noradrenaline are formed from dopamine. They act on the cardiac musculature and the metabolism (adrenaline) as well as on the peripheral circulation (noradrenaline) and help the body to cope with acute and chronic stress.

An increased production of catecholamines can be found with tumours of the chromaffine system (pheochromocytoma, neuroblastoma, ganglio­neuroma). An increased or decreased concentration of the catechol­amines can also be found with hypertension, degenerative cardiac diseases, schizophrenia and manic-depressive psychosis. The Noradrenaline (Norepinephrine) High Sensitive ELISA assay kit provides materials for the quantitative measurement of noradrenaline in low concentrated samples and for small sample volumes. Noradrenaline is extracted using a cis-diol-specific affinity gel and acylated to N-acylnoradrenaline and then converted enzymatically into N-acylnormetanephrine.

Assay Procedure

1. Pipette each 100 µl prepared Standards, Controls and Samples into the respective wells (colour coded yellow).
2. Pipette each 20 µl Noradrenaline-Antiserum (colour coded yellow) into all wells.
3. Cover the plate with adhesive foil, shake briefly and incubate for 15 – 20 hours (overnight) at 2 – 6 °C.
4. Discard or aspirate the contents of the wells and wash thoroughly with each 250 µl prepared Wash Buffer. Remove residual liquid by tapping the inverted plate on clean absorbent paper. Repeat the washing procedure 3 times.
5. Pipette each 100 µl POD-Conjugate into all wells.
6. Incubate for 60 minutes at room temperature on an orbital shaker (medium shaking rate).
7. Washing: Repeat wash step 4.Pipette each 100 µl Substrate into all wells.
   Incubate for 35 to 45 minutes at room temperature (20 – 25 °C) on an orbital shaker (medium shaking rate). Avoid exposure to direct sun light.
8. Pipette 100 µl Stop Solution into all wells.
9. Read the optical density at 450 nm (reference wavelength between 570 and 650 nm) in a microplate photometer within 15 minutes.

Typical Standard Curve