Quantitative Determination of Digoxin Concentration in Human Serum or Plasma by a Microplate Enzyme Immunoassay.





Materials Provided with Digoxin ELISA Test Kits:
1. Human Serum References: A-F
2. Dig Enzyme Reagent-HRP
3. Digoxin Biotin Reagent
4. Streptavidin Coated Plate
5. Wash Solution Concentrate
6. Substrate A: TMB
7. Substrate B: H202
8. Stop Solution

Materials & Instrumentations required, not included:
1. Precision pipettes
2. Distilled water
3. EIA kit Microplate Washer
4. EIA kit Microplate Reader at 450nm wavelength





Digoxin ELISA kit Background Information:
The clinical usefulness of the measurement of serum digoxin (DIG) is due to its low therapeutic ratio; a very small difference exists between therapeutic and toxic tissue levels. In addition, individuals may vary in their response to digoxin with an apparent increase in susceptibility to toxicity with age. The action of digoxin is to increase the force and velocity of myocardial contraction. This is necessary in the treatment of congestive heart failure and arrhythmias such as atrial fibrillation and atrial flutter. The myocardial concentrations of digoxin to serum levels remain relatively constant during normal renal function. This distribution ratio of digoxin is approximately 29 to 1 between the heart and serum. Thus, monitoring digoxin therapy by measurement of serum levels is feasible from the pharmacological standpoint, since serum levels are related to tissue levels following post-absorption equilibration. A practical and sensitive method of digoxin quantitation in serum is by enzyme immunoassay.

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Digoxin Elisa Test Principle:
This microplate enzyme immunoassay methodology provides the technician with optimum sensitivity while requiring few technical manipulations. In this method, serum reference, patient specimen, or control is first added to a microplate well. Enzyme-digoxin conjugate is added, then the reactants are mixed. A competition reaction results between the enzyme conjugate and the native digoxin for a limited number of antibody combining sites immobilized on the well. After the completion of the required incubation period, the antibody bound enzyme-digoxin conjugate is separated from the unbound enzyme-digoxin conjugate by aspiration or decantation. The activity of the enzyme present on the surface of the well is quantitated by reaction with a suitable substrate to produce color.





Digoxin ELISA Kits Performance and Characteristics:
A. Precision
The within and between assay precision of the DIG DAI ELISA test System were determined by
analyses on three different levels of pool control sera. The number (N), mean values (X), standard deviation (s) and coefficient of variation (C.V.) for each of these control sera are presented in Digoxin Elisa insert Table 2 and Table 3.
B. Sensitivity
The Digoxin procedure has an analytical sensitivity of 2.6 pg. This is equivalent to a sample containing a concentration of 0.05 ng/ml. The functional sensitivity (20%CV) was found to be 0.14 ng/ml.



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