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

INTENDED USE

3-CAT research kit is a procedure which can accurately measure in vitro quantitative levels of epinephrine, adrenaline, norepinephrine, noradrenaline, dopamine in EDTA-plasma, cell culture and many other biological sample types. This assay has a minimum sensitivity detection limit of 0.1 pg/ml for NAD and 0.25 pg/ml for AD and DOP.

BACKGROUND

Adrenaline, noradrenaline and dopamine are all synthesised in the; adrenal medulla the sympathetic nervous system and in the brain. These catecholamines are becoming increasing used as vital diagnostic markers for many diseases, since greater amounts of catecholamines and their metabolites (metanephrine and normetanephrine) are secreted in large quantities in these diseases. There has been a particular focus in tumor diseases that are linked with the nervous system such as; ganglioneuroma, pheochromocytoma and neuroblastoma.

This is based on a competitive procedure and the samples have all being been extracted using a cis-diol-specific affinity gel, acylated and then enzymatically derivatized. Since the chemical structure of catecholamines is identical in all animals and the fact that an extraction step has been performed this allows the kit to be used for a wide range of animal species.

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.

• Microtiter Plate – Empty in a resealable pouch.
• Adhesive Foil.
• Wash Buffer Concentrate (50x).
• Enzyme Conjugate.
• Substrate – TMB Chromogenic substrate.
• Stop Solution.
• Adrenaline Microtiter Strips – Antigen pre-coated microwell plate.
• Noradrenaline Microtiter Strips – Antigen pre-coated microwell plate.
• Dopamine Microtiter Strips – Antigen pre-coated microwell plate.
• Adrenaline Antiserum.
• Noradrenaline Antiserum.
• Dopamine Antiserum.
• Adjustment Buffer.
• TE Buffer.
• Standards (A-F) and Controls (1+2) – ADR: 0-80 ng/ml, NAD: 0-32 ng/ml, DOP: 0-80 ng/ml.
• Acylation Buffer.
• Coenzyme – S-adenosyl-L-methionine.
• Enzyme – Catechol-O-methyltransferase.
• Extraction Plate – Coated with boronate affinity gel.
• Hydrochloric Acid.

SENSITIVITY

The minimum detection levels of adrenaline, noradrenaline, epinephrine, norepinephrine, dopamine using current 3-CAT research kit was approximately 0.25 ng/ml for AD/DOP and 0.1 ng/ml NAD. The dynamic range for this assay is AD/DOP: 0.5 – 80 ng/ml and NAD: 0.2 – 32 ng/ml.

ASSAY CHARACTERISTICS

– Analytical Sensitivity Adrenaline Noradrenaline Dopamine (Limit of Detection): AD= 0.3 ng/mL x C*, NAD= 0.2 ng/mL x C* and DOP= 0.7 ng/mL x C* (C* = Correction factor).
– Intra-Assay: 9.3 – 17.1% (AD), 8.4 – 15.6% (NAD) and 10.8 – 21.5% (DOP).
– Inter-Assay: 11.2 – 16.3% (AD), 8.4 – 12.2% (NAD) and 14.8 – 23.0% (DOP).
– Recovery: AD= 12.6% (EDTA-Plasma), 8.7% (Cell Culture Medium), NAD= 6.9% (EDTA-Plasma), 17.7% (Cell Culture Medium) and DOP= 12.1% (EDTA-Plasma), 12.2% (Cell Culture Medium).

REFERENCES

1. Recent advances in methods for the analysis of catecholamines and their metabolites. Anal Bioanal Chem. (2006) 386 (3): 506-14. Review. Tsunoda M.
2. Identifying links in the chain: the dynamic coupling of catecholamines, peptide synthesis, and peptide release in hypothalamic neuroendocrine neurons. Adv Pharmacol. (2013) 68: 421-44. Review. Watts A.G. and Khan A.M.
3. Catecholamines and stress. Ideggyogy Sz. (2014) 67 (3-4): 116-20. Review. Palkovits M.
4. Recent insights on circulating catecholamines in hypertension. Curr Hypertens Rep. (2014) 16 (12): 498. Review. Ton Q.V. and Hammes S.R.
5. Neurotransmitters as regulators of tumor angiogenesis and immunity: the role of catecholamines. J Neuroimmune Pharmacol. (2013) 8 (1): 7-14. Review. Sarkar C., et al.
6. Identifying links in the chain: the dynamic coupling of catecholamines, peptide synthesis, and peptide release in hypothalamic neuroendocrine neurons. Adv Pharmacol. (2013) 68: 421-44. Review. Watts A.G. and Khan A.M.

ADDITIONAL INFORMATION

• Full Name: 3-CAT Research ELISA Kit (Adrenaline/Noradrenaline/Dopamine)
• Reactivity: Any Species
• Sample Type: EDTA-Plasma, Cell Culture, Other Biological Samples
• Sensitivity:  AD/DOP: 0.25 ng/ml and NAD: 0.1 ng/ml

OTHER RELATED ELISA KITS

• Adrenaline (Epinephrine) Research ELISA Kit
• Noradrenaline (Norepinerphrine) Research ELISA Kit
• Dopamine Research ELISA Kit