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Thyroglobulin ELISA 试剂盒

TG 适用: 人 Colorimetric Sandwich ELISA
产品编号 ABIN649039
发货至: 中国
  • 抗原 See all Thyroglobulin (TG) ELISA试剂盒
    Thyroglobulin (TG)
    适用
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    检测方法
    Colorimetric
    实验类型
    Sandwich ELISA
    应用范围
    ELISA
    原理
    Immunoenzymometric sequential assay (TYPE 4): The essential reagents required for an immunoenzymometric assay include high affinity and specificity antibodies (enzyme 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 Thyroglobulin antibody. Upon mixing monoclonal biotinylated antibody, and a serum containing the native antigen, reaction results between the native antigen and the antibody, forming an antibody-antigen complex. Simultaneously the biotin attached to the antibody binds to the streptavidin coated on the microwells resulting in immobilization of the complex. After a suitable incubation period, the antibody-antigen bound fraction is separated from unbound antigen by decantation or aspiration. Another antibody (directed at a different epitope) labeled with an enzyme is added. Another interaction occurs to form an enzyme labeled antibody-antigen-biotinylated-antibody complex on the surface of the wells. Excess enzyme is washed off via a wash step. A suitable substrate is added to produce color measurable with the use of a microplate spectrophotometer. The enzyme activity on the well is directly proportional to the native antigen concentration. By utilizing several different serum references of known antigen concentration, a dose response curve can be generated from which the antigen concentration of an unknown can be ascertained.
    Analytical Method
    Quantitative
    组件
    A. Thyroglobulin Calibrators (1. 0 ml/vial) A - FSix vials of references for Thyroglobulin antigen at levels of 0 (A), 2. 0 (B), 10. 0 (C), 40 (D), 100 (E), and 250 (F) ng/ml. A preservative has been added. Note: There is no known, internationally accepted thyroglobulin standard available. The Tg used in the serum based calibrators is a highly purified (98+% pure) human Tg preparation that is calibrated gravimetrically against the reference material obtained from Community Bureau of Reference Number CRM 457. B. x-Tg Biotin Reagent (13ml/vial): One vial containing biotinylated anti-Tg monoclonal mouse IgG in buffer, dye, and preservative. Store at 2-8°C. C. Tg Enzyme Reagent (13 ml/vial). One vial containing anti-thyroglobulin IgG labeled with horseradish peroxidase (HRP) in buffer, dye, and preservative. Store at 2-8°C. D. Streptavidin Coated Plate (96 wells). One 96-well (breakwell modules) microplate coated with streptavidin and packaged in an aluminum bag with a drying agent. Store at 2-8°C. E. Wash Solution Concentrate (20 ml). One vial containing a surfactant in buffered saline. A preservative has been added. Store at 2-30°C. F. Substrate A (7ml/vial). One bottle containing tetramethylbenzidine (TMB) in buffer. Store at 2-8°CG. Substrate B (7ml/vial). One bottle containing hydrogen peroxide (H2O2) in buffer. Store at 2-8°C. H. Stop Solution (8ml/vial). One bottle of stop solution containing a strong acid (1N HCl). Store at 2-8°C. I. Product Instructions: Note 1: Do not use reagents beyond the kit expiration date. Note 2: Opened reagents are stable for 60 days when stored at 2-8°C.
    试剂未包括
    1. Pipette (s) capable of delivering 50µl and 100µl 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% (optional). 3. Microplate washer or a squeeze bottle (optional). 4. Microplate Reader with 450nm and 620nm wavelength absorbance capability (The 620nm filter is optional). 5. Absorbent Paper for blotting the microplate wells. 6. Plastic wrap or microplate cover for incubation steps. 7. Vacuum aspirator (optional) for wash steps. 8. Timer. 9. Storage container for storage of wash buffer. 10. Distilled or deionized water. 11. Quality Control Materials.
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  • 应用备注
    Precautions: 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
    样本量
    50 μL
    板类型
    Pre-coated
    试剂准备
    1. Wash Buffer: Dilute contents of wash concentrate to 1000ml with distilled or deionized water in a suitable storage container. Store at room temperature (20-27°C) for up to 60 days. 2. Working Substrate Solution: Pour the contents of the amber vial labeled Solution A into the clear vial labeled Solution B. Place the yellow cap on the clear vial for easy identification. Mix and label accordingly. Store at 2 - 8 °C. Note: Do not use if the working substrate looks blue.
    样品收集
    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 without additives or gel barrier. Allow the blood to clot. Centrifuge the specimen to separate the serum from the cells. Samples may be refrigerated at 2-8°C for a maximum period of five days. If the specimen(s) cannot be assayed within this time, the sample(s) may be stored at temperatures of -20 °C for up to 30 days. Avoid repetitive freezing and thawing. When assayed in duplicate, 0. 100ml of the specimen is required.
    结果分析

    A dose response curve is used to ascertain the concentration of human thyroglobulin (Tg) in unknown specimens. 1. Record the absorbance obtained from the printout of the microplate reader. 2. Plot the absorbance for each duplicate serum reference versus the corresponding Tg concentration in ng/ml on linear graph paper (do not average the duplicates of the serum references before plotting). Draw the best-fit curve through the plotted points. 4. To determine the concentration of Tg for an unknown, locate the average absorbance of the duplicates for each unknown on the vertical axis of the graph, find the intersecting point on the curve, and read the concentration (in ng/ml) from the horizontal axis of the graph (the duplicates of the unknown may be averaged as indicated). Note: Computer data reduction software designed for IEMA (ELISA) assays may also be used for the data reduction.

    限制
    仅限研究用
  • 注意事项
    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 calibrator, control and patient sample 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 (50µl) of the appropriate calibrators, controls and samples into the assigned wells. 3. Add 0. 100 ml (100µl) of the x-Tg Biotin Reagent to each well. It is very important to dispense all reagents close to the bottom of the microwell. 4. Swirl the microplate gently for 20-30 seconds to mix. Cover with a plastic wrap or microplate cover. 5. Incubate for 16-20 hours 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 350µl of wash buffer (see Reagent Preparation Section), decant (tap and blot) or aspirate. Repeat two additional times for a total of three washes. An automatic or manual plate washer can be used. Follow the manufacturer's instruction for proper usage. If a squeeze bottle is used, fill each well to the top by squeezing the container (Avoiding air bubbles). Decant the wash and Repeat two additional times. Add 0. 100 ml (100µl) of Tg Enzyme Reagent to all wells DO NOT SHAKE THE PLATE AFTER ENZYME ADDITION. 9. Cover with a plastic wrap. Incubate at room temperature for 120 minutes. 10. Repeat steps 6 7. 11. Add 0. 100 ml (100 µl) of working substrate to all wells (See Reagent Preparation Section). Always add reagents in the same order to minimize reaction time differences between wells. DO NOT SHAKE THE PLATE AFTER SUBSTRATE ADDITION. 12. Cover with a plastic wrap or microplate cover. Incubate at room temperature for 15 minutes. 13. Add 0. 050ml (50µl) of stop solution to each well and mix gently for 15-20 seconds. Always add reagents in the same order to minimize reaction time differences between wells. 14. Read the absorbance in each well at 450nm (using a reference wavelength of 620-630nm to minimize well imperfections) in a microplate reader. The results should be read within 30 minutes of adding the stop solution. ALTERNATIVE SHORT PROCEDURE: This procedure can be used with the help of a laboratory hematology shaker. 1. Format the microplates' wells for each calibrator, control and patient sample 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 (50µl) of the appropriate calibrators, controls and samples into the assigned wells. 3. Add 0. 100 ml (100µl) of the biotin labeled monoclonal antibody to each well. It is very important to dispense all reagents close to the bottom of the microwell. 4. Incubate at room temperature for 2 hours while shaking constantly on a hematology shaker at 150 RPM. 5. Follow steps 6-14 as described in the Test Procedure above.
    储存条件
    4 °C/-20 °C
  • 抗原 See all Thyroglobulin (TG) ELISA试剂盒
    Thyroglobulin (TG)
    Abstract
    TG 产品
    别名
    TG ELISA Kit, cb717 ELISA Kit, AITD3 ELISA Kit, TGN ELISA Kit, Tgn ELISA Kit, cog ELISA Kit, thyroglobulin ELISA Kit, TG ELISA Kit, tg ELISA Kit, LOC701526 ELISA Kit, LOC701756 ELISA Kit, LOC100434297 ELISA Kit, Tg ELISA Kit
    背景
    Summary and Explanation of the test: Human thyroglobulin (Tg) is a large glycoprotein (660 kD) that is stored in the follicular colloid of the thyroid gland. It functions as a prohormone in the intrathyroid synthesis of primary thyroid hormones like Triiodothyronine (T3) and Thyroxine (T4). Tg is elevated in thyroid follicular and papillary carcinoma, thyroid adenoma, subacute thyroiditis, Hashimoto's thyroiditis and Graves Disease. Tg levels are found to be normal in patients with medullary thyroid carcinoma. Serial measurements of Tg is most useful in detecting recurrence of differentiated thyroid carcinoma following surgical resection or radioactive iodine ablation. Tg determination is used as an adjunct to iodine scanning but not as a replacement for it. Assessment of Tg levels aids in management of infants with congenital hypothyroidism. Tg determination has been done with various methods using direct competitive binding RIA and double antibody sandwich IRMA or Elisa, of which latter is more useful. All these methods suffer from interference by endogenous autoantibodies to Tg. It is useful to determine the effect of autoantibodies before screening such patients for levels of Tg. Intended Use: The Thyroglobulin Microplate Elisa test is intended to be used for the quantitative determination of thyroglobulin levels in human serum. The test is for in vitro diagnostic use only.
    途径
    Thyroid Hormone Synthesis
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