2-CAT Research ELISA Kit (Adrenaline/Noradrenaline)

INTENDED USE

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

BACKGROUND

Adrenaline and noradrenaline are both described as being catecholamines. Epinephrine acts on every body tissue but has varying effects depending on which tissue it is acting on. It is produced from norepinephrine from the adrenal medulla. It has been well documented that the synthesis of epinephrine is elevated during times of stress (physical and emotional).

Norepinephrine is produced from nerve axons (shaft of the nerve) and it is stored in small fluid-filled sacs which are called vesicles. During an action potential (usually an electrical pulse), it is released and can travel down the nerve and across the gap between two nerves where it binds to a receptor on the second nerve, this action stimulates the nerve to respond. This action is useful in causing vasoconstriction which is vital in either maintaining or increasing blood pressure.

This is a flexible competitive assay system that can be used for number of different biological sample types and volumes. The samples 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.

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

SENSITIVITY

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

ASSAY CHARACTERISTICS

– Intra Assay Precision: AD= 9.3 – 17.1% (EDTA-Plasma), 11.2 – 16.3% (RPMI) and NAD= 8.4 – 15.6% (EDTA-Plasma), 8.4 – 12.2% (RPMI).
– Recovery: AD= 12.1% (EDTA-Plasma), 8.7% (RPMI) and NAD= 6.9% (EDTA-Plasma), 17.7% (RPMI).

REFERENCES

Peripheral and central effects of circulating catecholamines. Compr Physiol. (2015) 5 (1): 1-15. Review. Tank A.W. and Lee Wong D.
Norepinephrine: more of a neurohormone than a vasopressor. Crit Care. (2010) 14 (5): 196. Review. Myburgh J.
Ficts and facts of epinephrine and norepinephrine stability in injectable solutions. Int J Pharm. (2012) 434 (1-2): 468-80. Review. Hoellein L. and Holzgrabe U.
Cardiac arrest: should we consider norepinephrine instead of epinephrine? Am J Emerg Med. (2014) 32 (12): 1560.e1-2. Mion G., et al.

2-CAT Research ELISA Kit (Adrenaline/Noradrenaline)

INTENDED USE

BACKGROUND

CONTENT

SENSITIVITY

ASSAY CHARACTERISTICS

REFERENCES

ADDITIONAL INFORMATION

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