Free Thyroxine Kit CLIA

N° du produit ABIN504745

Kit ELISA Free Thyroxine Humain, Chemiluminescent test pour la quantification de Humain Free Thyroxine.

Aperçu rapide pour Free Thyroxine Kit CLIA (ABIN504745)

Antigène: Free Thyroxine (fT4) (Free Thyroxine T4 (fT4))

Reactivité: Humain

Méthode de détection: Chemiluminescent

Type de méthode: Competition ELISA

Application: ELISA

Détail du produit Free Thyroxine Kit CLIA

Fonction: Competitive Chemiluminescence Immunoassay Analog Method for Free T4 (Type 5) The essential reagents required for a solid phase enzyme immunoassay include immobilized antibody, enzyme-antigen conjugate and native antigen. Upon mixing immobilized antibody, enzyme-antigen conjugate and a serum containing the native free antigen, a competition reaction results between the native free antigen and the enzyme-antigen conjugate for a limited number of insolubulized binding sites.

Analytical Method: Quantitative

Attributs du produit: The Quantitative Determination of Free Thyroxine Concentration in Human Serum by a Microplate Chemiluminescence immunoassay (CLIA)

Information d'application

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.

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. The blood should be collected in a plain redtop venipuncture tube without additives or gel barrier. Allow the blood to clot. Centrifuge the specimen to separate the serum from the cells. 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. When assayed in duplicate, 0.10ml 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. 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 microplate 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 0.050 ml (50l) of the appropriate serum reference, control or specimen into the assigned well. 3. Add 0.100 ml (100l) of fT4 Tracer Reagent to all wells. 4. Swirl the microplate gently for 20-30 seconds to mix and cover. 5. Incubate 45 minutes at room temperature. 6. Discard the contents of the microplate by decantation or aspiration. If decanting, 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 to all wells (see Reagent Preparation Section). ). Always add reagents in the same order to minimize reaction time differences between wells Incubate for five (5) minutes in the dark. 10. Read the relative light units in each well for 0.2 1.0 seconds. The results should be read within thirty (30) minutes of adding the signal solution.

Restrictions: For Research Use only

Stockage

Détail du antigène

Antigène: Free Thyroxine (fT4) (Free Thyroxine T4 (fT4))

Sujet: Thyroxine, the principal thyroid hormone, circulates in blood almost completely bound to carrier proteins. The main carrier is thyroxine-binding globulin (TBG). However, only the free (unbound) portion of thyroxine is responsible for the biological action. Further, the concentrations of the carrier proteins are altered in many clinical conditions, such as pregnancy. In normal thyroid function as the concentrations of the carrier proteins alters, the total thyroxine level changes so that the free thyroxine concentration remains constant. Thus, measurements of free thyroxine concentrations correlate better with clinical status than total thyroxine levels. The increase in total thyroxine associated with pregnancy, oral contraceptives and estrogen therapy occasionally result in total T4 levels over the limits of normal while the free thyroxine concentration remains in the normal reference range. Masking of abnormal thyroid function can also occur in both hyper and hypothyroid conditions by alterations in the TBG concentration. The total T4 can be elevated or lowered by TBG changes such that the normal reference levels result. The free thyroxine concentration can help in uncovering the patients actual clinical status. In this method, serum reference, patient specimen, or control is first added to a microplate well. Enzyme-T4 conjugate (analog method) is added and the reactants are mixed. A competition reaction results between the enzyme conjugate and the free thyroxine for a limited number of antibody combining sites immobilized on the well. After the completion of the required incubation period, the antibody bound enzyme-thyroxine conjugate is separated from the unbound enzyme-thyroxine conjugate via a wash step. 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 free thyroxine concentration permits construction of a graph of activity and con_x001F_centration. From comparison to the dose response curve, an unknown specimen's activity can be correlated with free thyroxine concentration.