Triiodothyronine T3 CLIA Kit
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北京 101111
Quick Overview for Triiodothyronine T3 CLIA Kit (ABIN504747)
抗原
See all Triiodothyronine T3 (T3) CLIA Kits适用
检测方法
实验类型
应用范围
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原理
- Competitive Chemiluminescence Immunoassay (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 antigen, a competition reaction results between the native antigen and the enzyme-antigen conjugate for a limited number of insolubilized binding sites.
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Analytical Method
- Quantitative
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产品特性
- The Quantitative Determination of Total Triiodothyronine Concentration in Human Serum or Plasma by a Microplate Chemiluminescence Immunoassay (CLIA)
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Triiodothyronine T3 (T3) ELISA Kit
T3 适用: Various Species Colorimetric Competition ELISA 123.5 pg/mL - 10000 pg/mL Plasma, Serum
Triiodothyronine T3 (T3) ELISA KitT3 适用: 大鼠 Colorimetric Competition ELISA 0.156 ng/mL - 10 ng/mL Plasma, Serum, Tissue Homogenate
Triiodothyronine T3 (T3) ELISA KitT3 适用: Various Species Colorimetric Competition ELISA 0.156 ng/mL - 10 ng/mL Plasma, Serum, Tissue Homogenate
Triiodothyronine T3 (T3) ELISA KitT3 适用: Pig Colorimetric Competition ELISA 0.156 ng/mL - 10 ng/mL Plasma, Serum, Tissue Homogenate
Triiodothyronine T3 (T3) ELISA KitT3 适用: Cow Colorimetric Competition ELISA 0.156 ng/mL - 10 ng/mL Plasma, Serum, Tissue Homogenate
Triiodothyronine T3 (T3) ELISA KitT3 适用: 小鸡 Colorimetric Competition ELISA 0.156 ng/mL - 10 ng/mL Plasma, Serum, Tissue Homogenate
Triiodothyronine T3 (T3) ELISA KitSmall Sample T3 适用: Various Species Colorimetric Competition ELISA 123.5 pg/mL - 10000 pg/mL Plasma, Serum
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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.
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板类型
- Pre-coated
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实验流程
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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 T3 procedure. Specimen(s) may be refrigerated at 2_x001E_8(C for a maximum period of 48 hours. If the specimen(s) can not be assayed within 48 hours, the sample(s) may be stored at temperatures of _x001E_20(C for up to 30 days. Before assay, allow the specimens to equilibrate to ambient temperature (20C 27C). When assayed in duplicate, 0.100ml of the specimen is required. . Materials [Required But Not Provided]: 1 Pipette capable of delivering 50l volumes with a precision of better than 1.5%. 2. Dispenser(s) for repetitive deliveries of 0.100ml and 0.350ml volumes with a precision of better than 1.5%. 3. Adjustable volume (20-200l) and (200-1000l) dispenser(s) for conjugate and substrate dilutions. 4. Microplate washers or a squeeze bottle (optional). Microplate Luminometer. Test tubes for dilution of enzyme conjugate and signal A and B. 7. Absorbent Paper for blotting the microplate wells. 8. Plastic wrap or microplate cover for incubation steps. 9. Vacuum aspirator (optional) for wash steps. 10. Timer. 11. Quality control materials.Reagent Preparation:
1. Working Tracer - T3-enzyme Conjugate Solution Dilute the T3-Tracer 1:11 with Total T3/T4 Tracer buffer in a clean container. For example, dilute 160l of conjugate with 1.6ml of buffer for 16 wells (A slight excess of solution is made). This reagent should be used within twenty-four hours for maximum performance of the assay. Store at 2-8(C. General Formula: Amount of Buffer required = Number of wells ( 0.1 Quantity of T3-Enzyme necessary = # of wells * 0.01 i.e. = 16 x 0.1 = 1.6ml for Total T3/T4 Conjugate Buffer 16 x 0.01 = 0.16ml (160l) for T3 enzyme conjugate 2. 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. 3. 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 0.050 ml (50l) of the appropriate serum reference, control or specimen into the assigned well. 3. Add 0.100 ml (100l) of Working Tracer, T3-enzyme conjugate solution to all wells (see Reagent Preparation Section). 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. 8. 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 five (5) minutes in the dark at room temperature 10. Read the relative light units in each well, for minimum 0.5 1.0 seconds, using a microplate luminometer. The results should be read within thirty (30) minutes of adding the signal solution. Note: For re-assaying specimens with concentrations greater than 7.5 ng/ml, pipette 25l of the specimen and 25l of the 0 serum reference into the sample well (this maintains a uniform protein concentration). Multiply the readout value by 2 to obtain the triiodothyronine concentration. -
限制
- 仅限研究用
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- Triiodothyronine T3 (T3)
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别名
- Triiodothyronine
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物质类
- Amino Acid
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背景
- Measurement of serum triiodothyronine concentration is generally re_x001F_garded as a valuable tool in the diagnosis of thyroid dysfunction. This importance has provided the impetus for the significant improvement in assay methodology that has occurred in the last two decades. The advent of monospecific antiserum and the discovery of blocking agents to the T3 binding serum proteins have enabled the development of procedurally simple radioimmunoassay (1,2). 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-T3 conjugate is added, and then the reactants are mixed. A competition reaction results between the enzyme conjugate and the native triiodothyronine for a limited number of antibody combining sites immobilized on the well. After the completion of the required incubation period, the antibody bound T3-enzyme conjugate is separated from the unbound T3-enzyme 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 triiodothyronine 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 T3 concentration.
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