Epinephrine/Adrenaline ELISA Kit, Cat# 1960-10
The Epinephrine/Adrenaline ELISA Kit is a sensitive competitive ELISA for the measurement of Epinephrine in human plasma or other species. For in
vitro research use only.
Epinephrine/Adrenaline (Plasma) ELISA Kit Features
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Epinephrine/Adrenaline standards A-F (0, 20, 60, 200, 800, 3200 pg/ml) & Controls 1-2
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Acylation buffer, Acyaltion reagent, Enzymes, and extraction protocol
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Typical Std. curve Epinephrine (pg/ml)
Highly specific Adrenaline IgG pre-coated, stabilized, ready-to-use 96-well strip plate, suitable for
multiple runs over 6-12 months.
100-600 ul EDTA-plasma samples are extracted and acylated using the supplied reagents and
acylation protocol. 100 ul of the processed samples are used in the ELISA in an overnight
assay.
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Sensitivity ~ 7 pg/ml;
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Storage (2-4ºC) , Shelf life : 12-18 months
Contains all necessary reagents. For in vitro research use only.
Assay Procedure: Allow all reagents to reach room temperature. Arrange and label required number of strips.
Adrenaline (epinephrine) is extracted from a plasma sample (100-600 ul) by using a cis-diol-specific affinity gel, acylated and then modified
enzymatically. The competitive ELISA kit uses the microtiter plate format. The antigen is bound to the solid phase of the microtiter plate. The derivatized
standards, controls and samples and the solid phase bound analytes compete for a fixed number of antiserum binding sites. After the system is in
equilibrium, free antigen and free antigen-antiserum complexes are removed by washing. The antibody bound to the solid phase is detected by an antirabbit IgG-peroxidase conjugate using TMB as a substrate. The reaction is monitored at 450 nm. Quantification of unknown samples is achieved by
comparing their absorbance with a reference curve prepared with known standard concentrations.
Step 1.
Step 2.
Step 3.
Step 4.
Step 5.
Pipet 100 ul each of calibrators, controls, and extracted samples.
Pipete 50 ul of Epinehrine antiserum incubate overnight (15-20 hrs) at 4oC.
Aspirate and wash 4X. Add100 ul of enzyme conjugate. Mix gently and incubate for 30 min at RT on a shaker.
Aspirate and wash 4X Add 100 ul of TMB Substrate solution to all wells, mix gently, and incubate at RT for 20 min.
Pipet 100 ul of stop solution into each well and mix gently (blue color turns yellow). Measure absorbance at 450 nm. Determine antibody
conc in each sample using the standards (results are expressed in units/ml).
Performance Characteristics
Calculation
Calculate the Net A450 values of the duplicate (deduct zero values). Plot the ADRENALINE concentration versus the net A450 and calculate the
samples values from the standard curve.
Expected values: <100 pg/ml
Crossreactivity/Interference
Derivatized Adrenaline 100%; Derivatized Noradrenaline 0.20%; Derivatized Dopamine < 0.0007%; Metanephrine 0.64%; Normetanephrine 0.0009%; 3Methoxytyramine < 0.0007%; 3-Methoxy-4-hydroxyphenylglycol 0.03%; Tyramine < 0.0007%; Phenylalanine, Caffeinic acid, L-Dopa, Homovanillic acid,
Tyrosine, 3-Methoxy-4-hydroxymandelic acid
Intra-assay (human EDTA plasma samples):
High 1329.3 ± 372.6 pg/ml; SD 124.2 pg/ml, CV9.3%
Medium 412.1 ± 129.6 pg/ml; SD 43.2 pg/ml; CV10.5%
Low 37.9 ± 19.5 pg/ml; 6.5 pg/ml; CV17.1%
Linearity
range 92 - 545 pg/mL; serial dilutions up to 1:8; 81 – 121%
Recovery: Human EDTA-Plasma 104.0 pg/ml (mean) 89.4 – 128.3 pg/ml (range) 13.1 pg/ml SD; CV 12.6%
Species reactivity: This kit is optimized for human plasma but it can be used for other species (rat, mouse, monkey, bovine, pig, etc).
General Information
Epinephrine (also known as adrenaline or adrenalin) is a hormone and a neurotransmitter. Epinephrine has many functions in the body, regulating
heart rate, blood vessel and air passage diameters, and metabolic shifts; epinephrine release is a crucial component of the fight-or-flight response of the
sympathetic nervous system. In chemical terms, epinephrine is one of a group of monoamines called the catecholamines. It is produced in some
neurons of the central nervous system, and in the chromaffin cells of the adrenal medulla from the amino acids phenylalanine and tyrosine. Adrenaline
is used to treat a number of conditions including: cardiac arrest, anaphylaxis, and superficial bleeding. It has been used historically for bronchospasm
and hypoglycemia, but newer treatments for these, such as salbutamol, a synthetic epinephrine derivative, and dextrose, respectively, are currently
preferred.
Adrenaline may be quantified in blood, plasma, or serum as a diagnostic aid, to monitor therapeutic administration, or to identify the causative agent in a
potential poisoning victim. Endogenous plasma adrenaline concentrations in resting adults are normally less than 10 ng/L, but may increase by 10-fold
during exercise and by 50-fold or more during times of stress. Pheochromocytoma patients often have plasma adrenaline levels of 1000-10,000 ng/L.
Parenteral administration of adrenaline to acute-care cardiac patients can produce plasma concentrations of 10,000 to 100,000 ng/L. Adrenaline is
available in an autoinjector delivery system. EpiPens, AuviQs, Anapens, and Twinjects all use adrenaline as their active ingredient. Twinjects contain a
second dose of adrenaline in a separate syringe and needle delivery system contained within the body of the autoinjector.
1960-10-Flr
130429A
Alpha Diagnostic Intl Inc., USA; [email protected];  (800) 786-5777; (210) 561-9515; www.4adi.com