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TSH Kit CLIA

TSH Reactivité: Humain Chemiluminescent Sandwich ELISA
N° du produit ABIN504766
  • Antigène Voir toutes TSH Kits CLIA
    TSH (Thyroid Stimulating Hormone (TSH))
    Reactivité
    • 6
    • 5
    • 3
    • 3
    • 3
    • 3
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    Humain
    Méthode de détection
    Chemiluminescent
    Type de méthode
    Sandwich ELISA
    Application
    ELISA
    Fonction
    Immunoenzymometric assay (Type 3) The essential reagents required for an immunoenzymometric assay include high affinity and specificity antibodies (enzyme conjugated and immobilized), with different and distinct epitope recognition, in excess, and native antigen. In this procedure, the immobilization takes place during the assay at the surface of a microplate well through the interaction of streptavidin coated on the well and exogenously added biotinylated monoclonal anti-TSH antibody. Upon mixing monoclonal biotinylated antibody, the enzyme-labeled antibody and a serum containing the native antigen, reaction results between the native antigen and the antibodies, without competition or stearic hindrance, to form a soluble sandwich complex.
    Analytical Method
    Quantitative
    Attributs du produit
    The Quantitative Determination of Thyrotropin Concentration in Human Serum by a Microplate) Chemiluminescence Immunoassay (CLIA)
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  • Indications d'application
    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.
    Volume d'échantillon
    50 μL
    Plaque
    Pre-coated
    Protocole

    Specimien Collection and Preparation:

    The specimens shall be blood serum in type and the usual precautions in the collection of venipuncture samples should be observed. For accurate comparison to established normal values, a fasting morning serum sample should be obtained. The blood should be collected in a plain red-top venipuncture tube with or without gel barrier. Allow the blood to clot. Centrifuge the specimen to separate the serum from the cells. Samples may be refrigerated at 2_x001E_8(C for a maximum period of five (5) days. If the specimen(s) can not be assayed within this time, 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, 0.100ml of the specimen is required. _x000E_

    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. 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 0.025 ml (25l) of the appropriate serum reference, control or specimen into the assigned well. 3. Add 0.100 ml (100l) of the TSH Tracer Reagent to each well. It is very important to dispense all reagents close to the bottom of the coated well. 4. Swirl the microplate gently for 20-30 seconds to mix and cover. 5. Incubate 30 minutes at room temperature. 6. Discard the contents of the microplate by decantation or aspiration. If decanting, tap and blot the plate dry with absorbent paper. 7. 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. 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. 9. Incubate for ten (10) minutes at room temperature in the dark. 10. Read the RLUs (Relative Light Units) in each well in a microplate luminometer for at least 0.2 seconds/ well. The results can be read within 30 minutes of adding the signal solution.
    Restrictions
    For Research Use only
  • Antigène Voir toutes TSH Kits CLIA
    TSH (Thyroid Stimulating Hormone (TSH))
    Autre désignation
    Thyrotropin (TSH) (TSH Produits)
    Synonymes
    CG7665 Kit CLIA, DLGR-1 Kit CLIA, DLGR1 Kit CLIA, Dmel\CG7665 Kit CLIA, FSH Kit CLIA, FSH-TSH Kit CLIA, Fsh Kit CLIA, Fsh/CG7665 Kit CLIA, LGR1 Kit CLIA, LH/CG receptor Kit CLIA, RH44949p Kit CLIA, TSH Kit CLIA, dLGR1 Kit CLIA, Leucine-rich repeat-containing G protein-coupled receptor 1 Kit CLIA, Lgr1 Kit CLIA
    Classe de substances
    Hormone
    Sujet
    Measurement of the serum concentration of thyrotropin (TSH), a glycoprotein with a molecular weight of 28,000 Daltons and secreted from the anterior pituitary, is generally regarded as the most sensitive indicator available for the diagnosis of primary and secondary (pituitary) hypothyroidism (1, 2). The structure of human TSH is similar to that of the pituitary and placental gonadotropins, consisting of an 89-amino acid a-subunit which is similar or identical between these hormones and a 115-amino acid -subunit, which apparently confers hormonal specificity. The production of the 2 subunits is separately regulated with apparent excess production of the a-subunit. The TSH molecule has a linear structure consisting of the protein core with carbohydrate side chains, the latter accounts for 16% of the molecular weight. Increase in serum concentrations of TSH, which is primarily responsible for the synthesis and release of thyroid hormones, is an early and sensitive indicator of decrease thyroid reserve and in conjunction with decreased thyroxine (T4) concentrations is diagnostic of primary hypothyroidism. The expected increase in TSH concentrations demonstrates the classical negative feedback system between the pituitary and thyroid glands. That is, primary thyroid gland failure reduces secretion of the thyroid hormones, which in turn stimulates the release of TSH from the pituitary. Additionally, TSH measurements are equally useful in differentiating secondary and tertiary (hypothalamic) hypothyroidism from the primary thyroid disease. TSH release from the pituitary is regulated by thyrotropin releasing factor (TRH), which is secreted by the hypothalamus, and by direct action of T4 and triiodothyronine (T3), the thyroid hormones, at the pituitary. Increase levels of T3 and T4 reduces the response of the pituitary to the stimulatory effects of TRH. In secondary and tertiary hypothyroidism, concentrations of T4 are usually low and TSH levels are generally low or normal. Either pituitary TSH deficiency (secondary hypothyroidism) or insufficiency of stimulation of the pituitary by TRH (tertiary hypothyroidism) causes this. The TRH stimulation test differentiates these conditions. In secondary hypothyroidism, TSH response to TRH is blunted while a normal or delayed response is obtained in tertiary hypothyroidism. Further, the advent of immunoenzymometric assays has provided the laboratory with sufficient sensitivity to enable the differentiating of hyperthyroidism from euthyroid population and extending the usefulness of TSH measurements. This method is a second-generation assay, which provides the means for discrimination in the hyperthyroid-euthyroid range. In this method, TSH calibrator, patient specimen or control is first added to a streptavidin coated well. Biotinylated monoclonal and enzyme labeled antibodies (Abs) are added and the reactants mixed. Reaction between the various TSH antibodies and native TSH forms a sandwich complex that binds with the streptavidin coated to the well. After the completion of the required incubation period, the antibody bound enzyme-thyrotropin conjugate is separated from the unbound enzyme-thyrotropin 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 thyrotropin levels permits construction of a dose response curve of activity and concentration. From comparison to the dose response curve, an unknown specimen's activity can be correlated with thyrotropin concentration.
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