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Digoxin CLIA Kit

DIG 适用: 人 Chemiluminescent Competition ELISA
产品编号 ABIN504797
发货至: 中国
  • 抗原 See all Digoxin (DIG) products
    Digoxin (DIG)
    适用
    检测方法
    Chemiluminescent
    实验类型
    Competition ELISA
    应用范围
    ELISA
    Analytical Method
    Quantitative
    产品特性
    The Quantitative Determination of Digoxin Concentration in Human Serum or Plasma by a Microplate Chemiluminescence Immunoassay (CLIA)
  • 应用备注
    All products that contain human serum have been found to be non-reactive for Hepatitis B Surface Antigen, HIV 1&2 and HCV Antibodies by FDA required tests. Since no known test can offer complete assurance that infectious agents are absent, all human serum products should be handled as potentially hazardous and capable of transmitting disease. Good laboratory procedures for handling blood products can be found in the Center for Disease Control / National Institute of Health, "Biosafety in Microbiological and Biomedical Laboratories," 2nd Edition, 1988, HHS Publication No. (CDC) 88-8395.
    样本量
    50 μL
    板类型
    Pre-coated
    实验流程

    Specimien Collection and Preparation:

    Collect sample(s) by venipuncture in ten (10) ml silicone evacuated tube(s) or evacuated tube(s) containing EDTA or heparin. The usual precautions in the collection of venipuncture samples should be observed. Separate the red blood cells by centrifugation. Use serum or plasma for the total DIG procedure. Specimen(s) may be refrigerated at 2_x001E_8(C for a maximum period of 48 hours. If the specimen(s) cannot be assayed within 48 hours, the sample(s) may be stored at temperatures of _x001E_20(C for up to 30 days. Avoid repetitive freezing and thawing. When assayed in duplicate, 0.050ml of the specimen is required. The cross_x001E_reactivity of the digoxin antibody to selected substances was evaluated by adding the interfering substance to a serum matrix at various concentrations. The cross-reactivity was calculated by deriving a ratio between dose of interfering substance to dose of digoxin needed to displace the same amount of tracer. Substance Cross Reactivity Digoxin 1.000 Digitoxin 0.019 Digitoxigenin 0.017 Lanatoside A 0.016 Ouabain 0.001 Spirnolactone 0.001 Prednisone 0.001 Pregnenolone 0.001 Digitoxose 0.001 Di-Acetyldigoxin, (-Methyldigoxin, (-Acetyldigoxin completely cross react in the assay.

    Reagent Preparation:

    1. Wash Buffer Dilute contents of wash solution to 1000ml with distilled or deionized water in a suitable storage container. Diluted buffer can be stored at room temperature (20-27(C) for up to 60 days. _x000E_2. Working Signal Reagent Solution - Store at 2 - 8(C. Determine the amount of reagent needed and prepare by mixing equal portions of Signal Reagent A and Signal Reagent B in a clean container. For example, add 1 ml of A and 1ml of B per two (2) eight well strips (A slight excess of solution is made). Discard the unused portion if not used within 36 hours after mixing. If complete utilization of the reagents is anticipated, within the above time constraint, pour the contents of Signal Reagent B into Signal Reagent A and label accordingly.

    Test Procedure:

    Before proceeding with the assay, bring all reagents, serum references and controls to room temperature (20 - 27(C). 1. Format the microplates wells for each serum reference, control and patient specimen to be assayed in duplicate. Replace any unused microwell strips back into the aluminum bag, seal and store at 2-8(C. 2. Pipette 0.025ml (25l) of the appropriate serum reference, control or specimen into the assigned well. 3. Add 0.050 ml (50l) of Digoxin Enzyme Reagent to all the wells 4. Swirl the microplate gently for 20-30 seconds to mix. 5. Add 0.050 ml (50l) Digoxin Biotin Reagent to all wells 6. Swirl the microplate gently for 20-30 seconds to mix. 7. Cover and incubate for 30 minutes at room temperature. 8. Discard the contents of the microplate by decantation or aspiration. If decanting, blot the plate dry with absorbent paper. 9. Add 350l of wash buffer (see Reagent Preparation Section), decant (tap and blot) or aspirate. Repeat two (2) additional times for a total of three (3) washes. An automatic or manual plate washer can be used. Follow the manufacturers instruction for proper usage. If a squeeze bottle is employed, fill each well by depressing the container (avoiding air bubbles) to dispense the wash. Decant the wash and repeat two (2) additional times. 10. Add 0.100 ml (100l) of working signal reagent solution to all wells (see Reagent Preparation Section). Always add reagents in the same order to minimize reaction time differences between wells. 11. Incubate at room temperature for five (5) minutes in the dark. 12. Read the relative light units in each well with a Chemiluminescence microplate reader for 0.5-1.0 seconds. The results should be read within 30 minutes after adding the working Signal Reagent. Note: For re-assaying specimens with concentrations greater than 4 ng/ml, pipet 12.5l of the specimen and 12.5l of the 0 serum reference into the sample well. Multiply the readout value by 2 to obtain the digoxin concentration.
    限制
    仅限研究用
  • 抗原 See all Digoxin (DIG) products
    Digoxin (DIG)
    Abstract
    DIG 产品
    物质类
    Chemical
    背景
    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 (1). 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 (2). 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 (3). 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 (1). A practical and sensitive method of digoxin quantitation in serum is by enzyme immunoassay. 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, and 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 light. The employment of several serum references of known digoxin concentration permits construction of a graph of activity and concentration. From comparison to the dose response curve, an unknown specimen's activity can be correlated with digoxin concentration. _x000E_
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