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

TPO Ab CLIA Kit

TPO Ab 适用: 人 Chemiluminescent Sandwich ELISA
产品编号 ABIN504792
发货至: 中国
  • 抗原 See all TPO Ab products
    TPO Ab (Anti-Thyroid-Peroxidase Antibody (TPO Ab))
    适用
    • 3
    • 3
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    检测方法
    Chemiluminescent
    实验类型
    Sandwich ELISA
    应用范围
    ELISA
    原理
    Antibodies to thyroid peroxidase have been shown to be characteristically present from patients with Hashimoto thyroiditis (95%), idiopathic myedema (90%) and Graves Disease (80%) 1. In fact 72% of patients positive for anti-TPO exhibit some degree of thyroid dysfunction2. This has lead to the clinical measurement becoming a valuable tool in the diagnosis of thyroid dysfunction. Measurements of antibodies to TPO have been done, in the past, by Passive Hemaglutination (PHA). PHA tests do not have the sensitivity of chemiluminescence immunoassay and are limited by subjective interpretation. This procedure, with the enhanced sensitivity of CLIA, permits the detectability of subclinical levels of antibodies to TPO. In addition, the results are quantitated by a luminometer, which eliminates subjective interpretation. Monobind's microplate chemiluminescence immunoassay methodology provides the technician with optimum sensitivity while requiring few technical manipulations. In this method, serum reference, diluted patient specimen, or control is first added to a microplate well. Biotinylated Thyroid Peroxidase Antigen (TPO) is added, and then the reactants are mixed. Reaction results between the autoantibodies to TPO and the biotinylated TPO to form an immune complex, which is deposited to the surface of streptavidin coated wells through the high affinity reaction of biotin and streptavidin. After the completion of the required incubation period, aspiration or decantation separates the reactants that are not attached to the wells. An enzyme anti-human IgG conjugate is then added to permit quantitation of reaction through interacting with human IgG of the immune complex. After washing, the enzyme activity is determined by reaction with substrate to produce light. The employment of several serum references of known antibody activity permits construction of a graph of enzyme and antibody activities. From comparison to the dose response curve, an unknown specimen's enzyme activity can be correlated with auto-immune antibody level.
    Analytical Method
    Quantitative
    产品特性
    The Quantitative Determination of Thyroid Peroxidase (TPO) Autoantibodies in Human Serum or Plasma by a Microplate Chemiluminescence Immunoassay (CLIA). Measurements of TPO autoantibodies may aid in the diagnosis of certain thyroid diseases such as Hashimotos and Graves as well as nontoxic goiter.
  • 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.
    说明

    Dilution of Sample: 1-100

    样本量
    25 μL
    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 anti-Tg 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. When assayed in duplicate, 0.100ml of the diluted specimen is required. Materials Required But Not Provided: 1. Pipet(s) capable of delivering 10l, & 25l volumes with a precision of <1.5%. 2. Dispenser(s) for repetitive deliveries of 0.100ml, 0.350ml and 1.0ml volumes with a precision of better than 1.5%. 3. Microplate washer (optional) or squeeze bottle to wash plates. 4. Microplate luminometer. 5. Test tubes for dilution of patient samples and signal reagents A and B. 6. Absorbent Paper for blotting the microplate wells. 7. Plastic wrap or microplate cover for incubation steps. 8. Vacuum aspirator (optional) for wash steps. 9. Timer.

    Reagent Preparation:

    1. Serum Diluent Dilute the serum diluent concentrate to 200ml in a suitable container with distilled or deionized water. Store at 2-8(C. 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. 4. Patient Sample Dilution (1/100) Dispense 0.010ml (10l) of each patient specimen into 1ml of serum diluent. Cover and vortex or mix thoroughly by inversion. Store at 2-8(C for up to forty-eight (48) hours

    Test Procedure:

    Before proceeding with the assay, bring all reagents, serum references and controls to room temperature (20-27C). 1. Assemble the microwell strips 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 diluted patient specimen into the assigned well. 3. Add 0.100 ml (100l) of TPO Biotinylated Conjugate Solution. 4. Swirl the microplate gently for 20-30 seconds to mix and cover. 5. Incubate 30 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 anti-TPO Tracer Reagent to all wells. Always add reagents in the same order to minimize reaction time differences between wells. 9. Incubate for thirty (30) minutes at room temperature. 10. Discard the contents of the microplate by decantation or aspiration. If decanting, blot the plate dry with absorbent paper. 11. 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. 13. Incubate at room temperature for five (5) minutes in the dark. 14. 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. Note: For re-assaying specimens with concentrations greater than 500 IU/ml, dilute the sample an additional 1:5 or 1:10 using the original diluted material. Multiply by the dilution factor to obtain the concentration of the specimen.
    限制
    仅限研究用
  • 抗原 See all TPO Ab products
    TPO Ab (Anti-Thyroid-Peroxidase Antibody (TPO Ab))
    别名
    Antibodies to thyroid peroxidase (TPO) (TPO Ab 产品)
    别名
    thyroid peroxidase CLIA Kit, Tpo CLIA Kit
    物质类
    Antibody, Antibody, Antibody
    背景
    Antibodies to thyroid peroxidase have been shown to be characteristically present from patients with Hashimoto thyroiditis (95%), idiopathic myedema (90%) and Graves Disease (80%) 1. In fact 72% of patients positive for anti-TPO exhibit some degree of thyroid dysfunction2. This has lead to the clinical measurement becoming a valuable tool in the diagnosis of thyroid dysfunction. Measurements of antibodies to TPO have been done, in the past, by Passive Hemaglutination (PHA). PHA tests do not have the sensitivity of chemiluminescence immunoassay and are limited by subjective interpretation. This procedure, with the enhanced sensitivity of CLIA, permits the detectability of subclinical levels of antibodies to TPO. In addition, the results are quantitated by a luminometer, which eliminates subjective interpretation. Monobind's microplate chemiluminescence immunoassay methodology provides the technician with optimum sensitivity while requiring few technical manipulations. In this method, serum reference, diluted patient specimen, or control is first added to a microplate well. Biotinylated Thyroid Peroxidase Antigen (TPO) is added, and then the reactants are mixed. Reaction results between the autoantibodies to TPO and the biotinylated TPO to form an immune complex, which is deposited to the surface of streptavidin coated wells through the high affinity reaction of biotin and streptavidin. After the completion of the required incubation period, aspiration or decantation separates the reactants that are not attached to the wells. An enzyme anti-human IgG conjugate is then added to permit quantitation of reaction through interacting with human IgG of the immune complex. After washing, the enzyme activity is determined by reaction with substrate to produce light. The employment of several serum references of known antibody activity permits construction of a graph of enzyme and antibody activities. From comparison to the dose response curve, an unknown specimen's enzyme activity can be correlated with auto-immune antibody level.
You are here:
客服