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Dehydroepiandrosterone Sulfate CLIA Kit

Reaktivität: Human Chemiluminescent Competition ELISA
Produktnummer ABIN504757
  • Target Alle Dehydroepiandrosterone Sulfate CLIA Kits anzeigen
    Dehydroepiandrosterone Sulfate
    Reaktivität
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    Human
    Nachweismethode
    Chemiluminescent
    Methodentyp
    Competition ELISA
    Applikation
    ELISA
    Verwendungszweck
    Competitive Enzyme Immunoassay (TYPE 7): The essential reagents required for an enzyme immunoassay include antibody, enzyme-antigen conjugate and native antigen. Upon mixing biotinylated antibody, enzyme-antigen conjugate and a serum containing the native antigen, a competition reaction results between the native antigen and the enzyme-antigen conjugate for a limited number of antibody binding sites.
    Analytische Methode
    Quantitative
    Produktmerkmale
    The Quantitative Determination of Dehydroepiandrosterone Sulfate Concentration (DHEA-S) in Human Serum or Plasma by a Microplate Chemiluminescence Immunoassay
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  • Applikationshinweise
    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.
    Plattentyp
    Pre-coated
    Protokoll

    Specimien Collection and Preparation:

    The specimens shall be blood, serum or heparanised plasma in type and taken with the usual precautions in the collection of venipuncture samples. The blood should be collected in a redtop (with or without gel additives) venipuncture tube or for plasma use evacuated tube(s) containing heparin. Allow the blood to clot for serum samples. Centrifuge the specimen to separate the serum or plasma from the cells. Samples may be refrigerated at 2_x001E_8oC for a maximum period of five (5) days. If the specimen(s) cannot be assayed within this time, the sample(s) may be stored at temperatures of _x001E_20oC for up to 30 days. Avoid repetitive freezing and thawing. When assayed in duplicate, 0.020ml of the specimen is required.

    Reagent Preparation:

    1. Wash Buffer Dilute contents of Wash Concentrate to 1000ml with distilled or deionized water in a suitable storage container. Store diluted buffer at room temperature 20-27(C for up to 60 days. 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 1 ml 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 0.010 ml (10 L) of the appropriate serum reference, control or specimen into the assigned well. 3. Add 0.050 ml (50l) of the DHEA-S Enzyme Tracer to all wells. 4. Swirl the microplate gently for 20-30 seconds to mix. 5. Add 0.050 ml (50l) of Anti- DHEA-S 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 four (4) additional times for a total of five (5) 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 four (4) 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: Dilute the samples suspected of concentrations higher than 8.0 g/ml 1:5 and 1:10 with DHEA-S 0 ug/ml calibrator or patient serum pools with a known low value for DHEA-S.
    Beschränkungen
    Nur für Forschungszwecke einsetzbar
  • Target Alle Dehydroepiandrosterone Sulfate CLIA Kits anzeigen
    Dehydroepiandrosterone Sulfate
    Andere Bezeichnung
    Dehydroepiandrosterone sulfate (DHEA-S) (Dehydroepiandrosterone Sulfate Produkte)
    Hintergrund
    Dehydroepiandrosterone sulfate (DHEA-S) is the major C19 steroid secreted by the adrenal cortex, and is a precursor in testosterone and estrogen biosynthesis. DHEA-S, the sulfate ester of DHEA, is derived from sulfated precursors and by enzymatic conversion of DHEA in adrenal and extradrenal tissues. Due to the presence of a 17-oxo [rather than hydroxyl] group, DHEA-S possesses relatively weak androgenic activity, which for unsulfated DHEA has been estimated at ~10% that of testosterone [1]. However, the bioactivity of DHEA-S may be increased by its relatively high serum concentrations, approximately 100 to 1000-fold higher than DHEA or testosterone, and its weak affinity for sex-hormone binding globulin [2]. The physiologic role of DHEA-S is not well-defined. Serum levels are relatively high in the fetus and neonate, low during childhood, and increase during puberty [3, 4]. Increased levels of DHEA-S during adrenarche may contribute to the development of secondary sexual hair. DHEA-S levels show a progressive decline after the third decade of life [5]. Unlike DHEA, DHEA-S levels do not show significant diurnal variation, show little day-to-day variation, are not responsive to acute corticotropin administration [4], and do not vary significantly during the normal menstrual cycle [2]. This may be due to the slower metabolic clearance rate of DHEA-S as compared to DHEA [6]. Measurement of serum DHEA-S is a useful marker of adrenal androgen synthesis. Abnormally low levels have been reported in hypoadrenalism [3], while elevated levels occur in several conditions, including virilizing adrenal adenoma and carcinoma [7], 21-hydroxylase and 3-hydroxysteroid dehydrogenase deficiencies [2,6] and some cases of female hirsutism [2]. Since very little DHEA-S is produced by the gonads [2, 3], measurement of DHEA-S may aid in the localization of the androgen source in virilizing conditions. Methods for measurement of DHEA-S include gas-liquid chromatography, double-isotope derivative techniques, competitive protein-binding assays, and radioimmunoassay. Although significant cross-reactivity occurs with DHEA, androstenedione and androsterone, the relative concentrations of these competing substances in most normal and pathologic samples predicts a minimal effect on assay performance. The Monobind DHEA-S CLIA kit uses a specific anti-DHEA-S antibody, and does not require prior sample extraction of serum or plasma. Cross-reactivity to other naturally occurring and structurally related steroids is low. The employment of several serum references of known DHEA-S concentration permits construction of a graph of activity (light) and con_x001F_centration. From comparison to the dose response curve, an unknown specimen's activity can be correlated with DHEA-S concentration.
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