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2-CAT (Adrenaline/Noradrenaline) Research ELISA Kit

INTENDED USE

2-CAT (adrenaline/noradrenaline) 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 (also called epinephrine) is mainly functioning as a hormone and is formed in the adrenal medulla, however, there are small amounts of it which act as neurotransmitter as they are produced in nerve fibres. Whereas, noradrenaline (also known as norepinephrine) is a most common neurotransmitter and is also a hormone. Both adrenaline and noradrenaline are catecholamines. Epinephrine is able to act on almost every body tissue but can have varying effects depending on which tissue it is acting on. It is mostly 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).

This epinephrine/norepinephrine research ELISA kit is an ideal tool for analysing various different types of diagnostic applications. Norepinephrine is mainly 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 epinephrine or norepinephrine and adrenaline or noradrenaline are both extracted by using a cis-diol-specific affinity gel, acylated and then converted enzymatically.

ADRENALINE/NORADRENALINE RESEARCH 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.

  • 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 epinephrine/norepinephrine 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

– Adrenaline/Noradrenaline (Epinephrine/Norepinephrine) Research: ELISA
– 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

  1. Peripheral and central effects of circulating catecholamines. Compr Physiol. (2015) 5 (1): 1-15. Review. Tank A.W. and Lee Wong D.
  2. Norepinephrine: more of a neurohormone than a vasopressor. Crit Care. (2010) 14 (5): 196. Review. Myburgh J.
  3. 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.
  4. Cardiac arrest: should we consider norepinephrine instead of epinephrine? Am J Emerg Med. (2014) 32 (12): 1560.e1-2. Mion G., et al.

ADDITIONAL INFORMATION

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

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