电话:
400-7060-959
传真:
+86 10 56315212-8813
电子邮件:
orders@antibodies-online.cn

Thyroxine T4 CLIA Kit

T4 适用: 人 Chemiluminescent Competition ELISA
产品编号 ABIN504751
发货至: 中国
  • 抗原 See all Thyroxine T4 (T4) CLIA Kits
    Thyroxine T4 (T4)
    适用
    • 5
    • 5
    • 4
    • 4
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    检测方法
    Chemiluminescent
    实验类型
    Competition ELISA
    应用范围
    ELISA
    原理
    Competitive Enzyme 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 insolubulized binding sites.
    Analytical Method
    Quantitative
    产品特性
    The Quantitative Determination of Total Thyroxine Concentration in Human Serum or Plasma by a Microplate Chemiluminescence Immunoassay (CLIA)
    Featured
    Discover our best selling T4 ELISA Kit
    Top Product
    Discover our top product T4 ELISA Kit
  • 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.
    板类型
    Pre-coated
    实验流程

    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 T4 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, 0.05ml of the specimen is required. Materials Required [But Not Provided]: 1. Pipette capable of delivering 25l 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 washer or a squeeze bottle (optional). 5. Microplate Luminometer 6. 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 T4 Tracer Reagent Dilute the T4-Tracer 1:11 with Total T3/T4 tracer buffer in a suitable 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 T4 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 T4 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 0.025 ml (25l) of the appropriate serum reference, control or specimen into the assigned well. 3. Add 0.100 ml (100l) of Working T4 Tracer Reagent to all wells (see Reagent Preparation Section). 4. Swirl the microplate gently for 20-30 seconds to mix and cover. _x000E_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 at room temperature for five (5) minutes in the dark. 10. 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 signal substrate. Note: For reassaying specimens with concentrations greater than 25 g/dl, pipet 12.5l of the specimen and 12.5l of the 0 serum reference into the sample well (this maintains a uniform protein con_x001F_centration). Multiply the readout value by 2 to obtain the thyroxine concentration.
    限制
    仅限研究用
  • 抗原 See all Thyroxine T4 (T4) CLIA Kits
    Thyroxine T4 (T4)
    别名
    Thyroxine (T4 产品)
    物质类
    Amino Acid
    背景
    Measurement of serum thyroxine concentration is generally re_x001F_garded as an important in-vitro diagnostic test for assessing thyroid function. This importance has provided the impetus for the significant improvement in assay methodology that has occurred in the last three decades. This procedural evolution can be traced from the empirical protein bound iodine (PBI) test (1) to the theoretically sophisticated radioimmunoassay (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-T4 conjugate is added, and then the reactants are mixed. A competition reaction results between the enzyme conjugate and the native 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 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 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 thyroxine concentration.
You are here:
客服