3-CAT ELISA

Background:In humans the catecholamines adrenaline (epinephrine), noradrenaline (norepinephrine) and dopamine are neurotransmitters of the sympathetic nervous system and are involved in many physiological processes. The sympathetic nervous system sets the body to a heightened state of alert, also called as the body’s fight-or-flight response. In the human body the catecholamines and their metabolites indicate the adaptation of the body to acute and chronic stress. Next to the metanephrine/normetanephrine the catecholamines are important for the diagnosis and the follow-up of tumors of the sympathoadrenal system like the pheochromocytoma. The quantitative determination of catecholamines in urine is preferred for the diagnosis of these tumors, whereas the determination of catecholamines in plasma is medically sensible for the localization of the tumor and for function testing. Values above the cut-off can provide an indication for neuroendocrine tumors. However, in literature various diseases like hypertension, cardiovascular diseases, schizophrenia and manic depression are described with abnormal low or high levels of catecholamines.

Description:
Enzyme Immunoassay for the quantitative determination of adrenaline (epinephrine), noradrenaline (norepinephrine) and dopamine in plasma and urine. Adrenaline (epinephrine), noradrenaline (norepinephrine) and dopamine are extracted by using a cisdiol-specific affinity gel, acylated and then converted enzymatically. The subsequent competitive ELISA uses the microtiter plate format. The antigen is bound to the solid phase of the microtiter plate. The acylated standards, controls and samples compete with the solid phase bound analytes for a fixed number of antibody binding sites. After the system is in equilibrium, free antigen and free antigen-antibody complexes are removed by washing. The antibody bound to the solid phase is detected by an anti-rabbit IgG-peroxidase conjugate using TMB as a substrate resulting in a colour reaction. The reaction is monitored at a wavelength of 450 nm. Quantification of unknown samples is achieved by comparing their absorbance with a reference curve prepared with known standard concentrations. Manual processing of the ELISA is recommended. The use of automatic laboratory equipment is the responsibility of the user. This in-vitro diagnostic is for professional use only.

Product features:
The kit contains reagents for 1x96 determinations for adrenaline, 1x96 determinations for noradrenaline and 1x96 determinations for dopamine;
Microtiter plates consisting of 12x8;

Microtiter plate reader at 450 nm and, if possible, 620-650 nm;
Analytical sensitivity:
- AD: urine: 0.9 ng/ml; plasma: 10 pg/ml;
- NAD: urine: 1.7 ng/ml; plasma: 36 pg/ml; - DOP: urine: 2.5 ng/ml; plasma: 49 pg/ml;
IVD