SERPINA7 CLIA Kit

Produktnummer ABIN504776

Produktdetails SERPINA7 CLIA Kit

Target: SERPINA7 (Serpin Family A Member 7 (SERPINA7))

Reaktivität: Human

Nachweismethode: Chemiluminescent

Methodentyp: Sandwich ELISA

Applikation: ELISA

Verwendungszweck: Immunoenzymometric assay: The essential reagents required for an enzyme immunoassay include high affinity and specificity antibody in, enzyme labeled antigen and the 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 polyclonal anti-TBG antibody. Upon mixing polyclonal biotinylated antibody with the enzyme-labeled antigen and a serum containing the native antigen, a competition reaction results between the native antigen and the enzyme labeled antigen for a limited number of specific binding sites on the antibody. The antigen bound antibody attaches to the surface of the plastic wells because of the biotin label on it and the streptavidin that is present on the plastic well.

Analytische Methode: Quantitative

Produktmerkmale: The Quantitative Determination of TBG Thyroxine Binding Globulin concentration in Human Serum, Plasma or Whole Blood by a Microplate Enzyme Labeled Chemiluminescense Immunoassay

Anwendungsinformationen

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. For accurate comparison to establish normal values, a fasting morning serum sample should be obtained. The blood should be collected in a plain redtop venipuncture tube without additives or anti-coagulants. Allow the blood to clot for samples. 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) cannot 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, 0.020ml of the specimen is re_x001F_quired.

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 0.010 ml (10 l) of the appropriate serum reference, control or specimen into the assigned wells. 3. Add 0.050 ml (50 l) of the TBG Tracer Reagent to each well. Mix well the contents of the microwells. It is very important to dispense all reagents close to the bottom of the coated well. 4. Add 0.050 ml (50 l) of the TBG Biotin Reagent to each well. 5. Swirl the microplate gently for 20-30 seconds to mix and cover. 6. Incubate 30 minutes at room temperature. Discard the contents of the microplate by decantation or aspiration. If decanting, tap and blot the plate dry with absorbent paper. 8. 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. 9. Add 0.100 ml (100l) of working Signal Reagent to all wells (see Reagent Preparation Section). 10. Incubate for five (5) minutes at room temperature in the dark. 11. 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 substrate solution.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Antigendetails

Target: SERPINA7 (Serpin Family A Member 7 (SERPINA7))

Andere Bezeichnung: Thyroxine Binding Globulin (SERPINA7 Produkte)

Synonyme: C730040N12Rik CLIA Kit, Tbg CLIA Kit, TBG CLIA Kit, THYROXINE-BINDING GLOBULIN CLIA Kit, serine (or cysteine) peptidase inhibitor, clade A (alpha-1 antiproteinase, antitrypsin), member 7 CLIA Kit, serpin family A member 7 CLIA Kit, Serpina7 CLIA Kit, SERPINA7 CLIA Kit

Hintergrund: TBG (Thyroxine Binding Globulin) a 54 kD liver glycoprotein is the principal binding protein for T4 and T3 in circulation. Electrophoretic analyses indicate that T4 is bound, in decreasing order, to TBG, to a T4 binding prealbumin (TBPA) and to albumin. By virtue of its intense affinity for T4, TBG is by far the major determinant of overall binding capacity. The interaction between T4 and its binding proteins conforms to a reversible binding equilibrium in which the majority of the hormone is bound and a very small portion (< 0.05%) is free. T3 is not bound by TBPA and is bound by TBG less firmly than is T4. As a consequence proportion of free T3 is normally 8-10 times greater than T4. Only free (T3/T4) hormones are available to the tissues, therefore the metabolic state of the patient will correlate more closely with the free than with the total concentration of the hormones. The diagnostic accuracy of the total hormone measurements would be equal to the free hormone if all the patients had similar binding protein concentrations. Unfortunately, serum TBG abnormalities that distort the total:free relationship, are commonly encountered in clinical practice. Additionally the presence of antibodies to thyroid hormones, in some patients, renders total hormone measurements unreliable. Considerable confusion still exists regarding the validity of free hormone testing. There is controversy regarding the clinical utility of free hormone testing in conditions associated with binding protein abnormalities of pregnancy and non-thyroidal illness. Methods that are sensitive to albumin concentrations, the effect of certain drugs, high free fatty acid and levels of hormones binding inhibitors are considered inadequate by some researchers. However, the techniques for physically separating the exceedingly small amounts of free hormones from the dominant protein bound moiety are too technically demanding, inconvenient and expensive for a routine clinical laboratory. Such methods that employ equilibrium dialysis, ultrafilteration and gel-filtration are typically used by researchers. In routine analysis the clinical laboratories rely on direct measurements of free and total hormones and their binding proteins, mainly TBG. Based on their serum concentrations, familial TBG variants are divided into four major categories: excess, normal, partial deficiency and complete absence. The studies show that estrogens pregnancy and oral contraceptives acute intermittent porphyria and chronic liver disease increase TBG concentrations while, androgenic and anabolic steroids, large doses of glucocorticoids and nephrosis decreases TBG levels. In this method, TBG calibrator, patient specimen or control is first added to a streptavidin coated well. Biotinylated polyclonal antibody (highly specific for TBG) and enzyme labeled TBG are added, in sequence, and the reactants mixed. Reaction between the TBG antibodies, enzyme labeled TBG and native TBG forms a complex that binds with the streptavidin coated to the well. After the completion of the required incubation period, the excess enzyme conjugate is separated from the bound fraction 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 TBG 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 TBG concentration.

Pathways: Hormone Transport