2-CAT Research ELISA Kit (Noradrenaline/Dopamine)

INTENDED USE

2-CAT research ELISA kit is a reliable and accurate in vitro quantitative procedure for analysing noradrenaline, norepinephrine, dopamine in plasma (EDTA), cell culture and many other biological samples types. This assay has a minimum sensitivity detection limit of 0.1 ng/ml (NAD) and 0.25 ng/ml (DOP).

BACKGROUND

Noradrenaline (also called norepinephrine) is a neurotransmitter of the sympathetic nervous system that is responsible for reflexive and tonic changes in the cardiovascular tone. It must be noted that the sympathetic noradrenergic system is active all the time even when individuals are in a state of rest, it is working in order to maintain the tonic levels of cardiovascular performance. Naturally occurring norepinephrine are usually found to be made inside the shaft of nerve called nerve axons, they are then stored inside vesicles which are small fluid-filled sacs. They get released during an electrical impulse (action potential), this causes them to travel down the nerve, across the gap between two nerves and bind to a receptor on the second nerve, resulting in stimulating that nerve to respond. It acts mostly on alpha receptors, there have been a few examples where it stimulates beta receptors.

Dopamine is a neurotransmitter that helps control the brain’s pleasure centres and reward. It is the chemical that is found to mediate pleasure within the brain. It is always released during pleasurable situations or it can stimulate one to seek out the pleasurable occupation and activity. Examples include food, sleep, memory, mood, learning, many drugs of abuse and sex. It can help in focus and attention, vision is able to aid a dopamine-response in the brain and this can help one to direct their attention and focus. It is also thought to be responsible for short term memory that is based on an imagined response to certain information.

This is a flexible competitive assay system that is designed for many different biological sample types and volumes. The noradrenaline/norepinephrine and dopamine are both extracted by using a cis-diol-specific affinity gel, acylated and then converted enzymatically.

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 – 1 x 96 wells, empty in a resealable pouch.
• Adhesive Foil – Adhesive Foils in a resealable pouch.
• Wash Buffer Concentrate (50x).
• Enzyme Conjugate – Goat anti-rabbit immunoglobulins, conjugated with peroxidase.
• Substrate – Chromogenic substrate containing tetramethylbenzidine (TMB).
• Stop Solution – 0.25 M sulfuric acid.
• Noradrenaline Microtiter Strips – 1 x 96 well (12×8) antigen precoated microwell plate.
• Dopamine Microtiter Strips – 1 x 96 well (12×8) antigen precoated microwell plate.
• Noradrenaline Antiserum – Rabbit anti noradrenaline antibody.
• Dopamine Antiserum – Rabbit anti dopamine antibody.
• Adjustment TRIS Buffer.
• TE Buffer – TRIS-EDTA buffer.
• Extraction Plate – 2 x 48 well plates coated with boronate affinity gel.
• Hydrochloric Acid – 0.025 M Hydrochloric Acid.
• Standards (A-F) and Controls (1+2) – Concentration NAD: 0-32 ng/ml and DOP: 0-80 ng/ml.
• Acylation Buffer.
• Acylation Reagent.
• Coenzyme – S-adenosyl-L-methionine.
• Enzyme – Catechol-O-methyltransferase.

SENSITIVITY

The minimum sensitivity detection limit of noradrenaline/norepinephrine and dopamine using current 2-CAT research ELISA kit was approximately 0.1 ng/ml (NAD) and 0.25 ng/ml (DOP). The dynamic range for this assay is NAD: 0.2 – 32 ng/ml and DOP: 0.5-80 ng/ml.

ASSAY CHARACTERISTICS

– Intra Assay Precision: DOP= 10.8 – 21.5% (EDTA-Plasma), 14.8 – 23.0% (RPMI) and NAD= 8.4 – 15.6% (EDTA-Plasma), 8.4 – 12.2% (RPMI).
– Recovery: DOP= 12.1% (EDTA-Plasma), 12.2% (RPMI) and NAD= 6.9% (EDTA-Plasma), 17.7% (RPMI).

REFERENCES

1. Neurotransmitters of the brain: serotonin, noradrenaline (norepinephrine), and dopamine J Neurosci Nurs. (2000) 32 (4): 234-8. Review. Blows WT.
2. Dopamine versus norepinephrine: is one better? Minerva Anestesiol. (2009) 75 (5): 333-7. Review. Vincent J.L., et al.
3. Norepinephrine and dopamine as learning signals. Neural Plast. (2004) 11 (3-4): 191-204. Review. Harley C.W.
4. 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.
5. Dopamine reuptake by norepinephrine neurons: exception or rule? Crit Rev Neurobiol. (2004) 16 (1-2): 121-8. Review. Carboni E. and Silvagni A.

ADDITIONAL INFORMATION

• Full Name: 2-CAT Research ELISA Kit (Noradrenaline/Dopamine)
• Reactivity: Any Species
• Sample Type: EDTA-Plasma, Cell Culture, Other Biological Samples
• Sensitivity:  NAD: 0.1 ng/ml and DOP: 0.25 ng/ml

OTHER RELATED ELISA KITS

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