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2-CAT ELISA Kit (Adrenaline/Noradrenaline)

INTENDED USE

2-CAT ELISA kit is a reliable and accurate in vitro quantitative procedure for analysing epinephrine, norepinephrine, adrenaline, noradrenaline in plasma and urine samples. This assay has a minimum sensitivity detection limit of 10 pg/ml (P) or 0.3 ng/ml (U) for AD and 50 pg/ml (P) or 1.5 ng/ml (U) for NAD.

BACKGROUND

Adrenaline (epinephrine), is produced in the adrenal medulla, it is also made in small amounts in the nerve fibres. In general, naturally occurring norepinephrine is predominately produced inside nerve axons (usually in the shaft of the nerve) and then stored inside structure that are called vesicles (these are small fluid-filled sacs) until they are released when an action potential (in most cases an electrical pulse) travels down the nerve.

The synthesis of adrenaline increases during times of stress, where our adrenal medulla is helping us to cope with emotional or physical stress. One of the main function is to cause vasoconstriction (a process resulting in the narrowing of blood vessels), This is useful for either maintaining or even increasing blood pressure (during time of stress).

This is a competitive assay where the samples has been extracted through a cis-diol-specific affinity gel, acylated and then derivatized 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: 0-100 ng/ml and NAD: 0-1000 ng/ml.
  • Adhesive Foil – Adhesive Foils in a resealable pouch.
  • Wash Buffer Concentrate (50x) – Buffer with a non-ionic detergent and physiological pH.
  • Enzyme Conjugate – Goat anti-rabbit immunoglobulins, conjugated with peroxidase.
  • Substrate – Chromogenic substrate containing tetramethylbenzidine, substrate buffer and hydrogen peroxide.
  • Stop Solution – 0.25 M sulfuric acid.
  • Acylation Buffer.
  • Acylation Reagent.
  • Assay Buffer – 1M hydrochloric acid.
  • 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 ELISA kit was approximately 50 pg/ml (P) or 1.5 ng/ml (U) for NAD and 10 pg/ml (P) or 0.3 ng/ml (U) for AD. The dynamic range for this assay is AD: 1 – 200 ng/ml and NAD: 5 – 1,000 ng/ml.

ASSAY CHARACTERISTICS

– Intra Assay Precision: 6.9 – 15.8% (AD) and 9.8 – 16.1% (NAD)
– Inter Assay Precision: 13.2 – 18.2% (AD) and 8.5 – 15.0% (NAD)
– Linearity: 88 – 122% (AD) and 87 – 120.0% (NAD)
– Recovery: 88 – 122% (AD) and 87 – 120.0% (NAD)

REFERENCES

  1. Presynaptic metabotropic receptors for acetylcholine and adrenaline/noradrenaline. Handb Exp Pharmacol. (2008) (184): 261-88. Review. Gilsbach R. and Hein L.
  2. The effects of epinephrine/norepinephrine on end-tidal carbon dioxide concentration, coronary perfusion pressure and pulmonary arterial blood flow during cardiopulmonary resuscitation. Resuscitation. (2000) 43 (2): 129-40. Lindberg L., et al.
  3. Training and 24-hr urinary catecholamine excretion. Int J Sports Med. (2009) 30 (1): 33-9. Filaire E., et al.
  4. Individual differences in adrenaline/noradrenaline reactivity and self-perceived health status. Z Gesamte Hyg. (1990) 36 (8): 413-4. Mejman T.F., et al.

ADDITIONAL INFORMATION

  • Full Name: 2-CAT ELISA Kit (Adrenaline/Noradrenaline)
  • 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)

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