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3-CAT (Adrenaline/Noradrenaline/Dopamine) ELISA Kit

INTENDED USE

3-CAT (adrenaline/noradrenaline/dopamine) ELISA kit is an accurate and reliable in vitro quantitative method for detecting epinephrine, adrenaline, norepinephrine, noradrenaline, dopamine in plasma and urine. This assay has a minimum sensitivity detection limit of 0.01 pg/ml (P) or 0.8 ng/ml (U) for AD, 0.04 pg/ml (P) or 1.7 ng/ml (U) for NAD and 1.9 pg/ml (P) or 2.5 ng/ml (U) for DOP.

BACKGROUND

Adrenaline, noradrenaline and dopamine are all catecholamines which are synthesised in the adrenal medulla, the sympathetic nervous system and in the brain. They are found to known to effect virtually every tissue in the human body and can also work together with other neuronal and hormonal systems in order to regulate a wide range of physiological processes. Catecholamines and their metabolites (normetanephrine and metanephrine) have become useful markers for diagnostic purposes since they are secreted in greater amounts in a large number of diseases. In particular tumor diseases which are associated with the nervous system are of primary importance, examples include neuroblastoma, pheochromocytoma and ganglioneuroma.

This is a competitive assay where adrenaline, epinephrine, noradrenaline, norepinephrine, dopamine have been extracted using a cis-diol-specific affinity gel, acylated and then derivatized enzymatically. Due to the fact that an extraction step has been performed, this allows the kit to be used for a large number of animal species since the chemical structure of these catecholamines is identical in all animals.

ADRENALINE/NORADRENALINE/DOPAMINE ELISA KIT CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

  • Adhesive Foil.
  • Wash Buffer Concentrate (50x).
  • Enzyme Conjugate.
  • Substrate (Tetramethylbenzidine).
  • Stop Solution.
  • Adrenaline Microtiter Strips.
  • Noradrenaline Microtiter Strips.
  • Dopamine Microtiter Strips.
  • Antiserum – Rabbit anti-adrenaline antibody.
  • Antiserum – Rabbit anti-noradrenaline antibody.
  • Antiserum – Rabbit anti-dopamine antibody.
  • Adjustment Buffer.
  • Acylation Buffer.
  • Acylation Reagent.
  • Assay Buffer.
  • Enzyme – Catechol-O-methyltransferase.
  • Extraction Buffer.
  • Extraction Plate – 2 x 48 well plates coated with boronate affinity gel.
  • Hydrochloric Acid.
  • Standards (A-F) and Controls (1+2) – ADR: 0-200 ng/ml, NAD: 0-1,000 ng/ml and DOP: 0-2,000 ng/ml.

SENSITIVITY

The minimum sensitivity detection limit of adrenaline, epinephrine, noradrenaline, norepinephrine, dopamine using current epinephrine/norepinephrine/dopamine ELISA kit was approximately 0.3 ng/ml (U) or 10 pg/ml (P) for AD, 1.5 ng/ml (U) or 50 pg/ml (P) for NAD, and 2.5 ng/ml (U) or 1.9 pg/ml (P) for DOP. The dynamic range for this assay is AD: 1 – 200 ng/ml, NAD: 5 – 1,000 ng/ml and DOP: 10 – 2,000 ng/ml.

ASSAY CHARACTERISTICS

– Adrenaline/Noradrenaline/Dopamine (Epinephrine/Norepinephrine/Dopamine): ELISA
– Expected Reference Values: AD= < 20 µg/day (Urine), < 100 pg/mL (Plasma); NAD= < 90 µg/day (Urine), < 600 pg/mL (Plasma) and DOP= < 600 µg/day (Urine), < 100 pg/mL (Plasma).
– Analytical Sensitivity (Limit of Detection): AD= 0.3 ng/mL (Urine), 10 pg/mL (Plasma); NAD= 1.5 ng/mL (Urine), 50 pg/mL (Plasma) and DOP= 4.5 ng/mL (Urine), 25 pg/mL (Plasma).
– Intra Assay Precision: 6.9 – 15.0% (AD), 9.8 – 16.1% (NAD) and 9.5 – 15.8% (DOP).
– Inter Assay Precision: 13.2 – 15.4% (AD), 8.5 – 15.0% (NAD) and 15.9 – 18.2% (DOP).
– Linearity: 88 – 122% (AD), 87 – 120% (NAD) and 86 – 104% (DOP).
– Recovery: AD= 84 – 119% (Urine), 80 – 113% (Plasma), NAD= 83 – 115% (Urine), 85 – 108% (Plasma) and DOP= 85 – 104% (Urine), 74 – 110% (Plasma).

REFERENCES

  1. A new approach to reducing postsurgical cancer recurrence: perioperative targeting of catecholamines and prostaglandins. Clin Cancer Res. (2012) 18 (18): 4895-902. Review. Neeman E, et al.
  2. Norepinephrine and dopamine as learning signals. Neural Plast. (2004) 11 (3-4): 191-204. Review. Harley CW.
  3. Relevance of norepinephrine-dopamine interactions in the treatment of major depressive disorder. CNS Neurosci Ther. (2010) 16 (3): e1-17. Review. El Mansari M, et al.
  4. Determination of catecholamines in plasma and urine. Best Pract Res Clin Endocrinol Metab. (2013) 27 (5): 713-23. Review. Grouzmann E. and Lamine F.

ADDITIONAL INFORMATION

  • Full Name: 3-CAT (Adrenaline/Noradrenaline/Dopamine) ELISA Kit
  • Reactivity: Any Species
  • Sample Type: Plasma, Urine
  • Sensitivity:  AD: 0.3 ng/ml (U); 10 pg/ml (P) and NAD: 1.5 ng/ml (U); 50 pg/ml (P) and DOP: 2.5 ng/ml (U); 1.9 pg/ml (P)

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