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alpha Fetoprotein CLIA Kit

AFP 适用: 人 Chemiluminescent Sandwich ELISA
产品编号 ABIN504777
发货至: 中国
  • 抗原 See all alpha Fetoprotein (AFP) CLIA Kits
    alpha Fetoprotein (AFP) (alpha-Fetoprotein (AFP))
    适用
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    检测方法
    Chemiluminescent
    实验类型
    Sandwich ELISA
    应用范围
    ELISA
    原理
    Immunoenzymometric assay: 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 anti-AFP antibody. Upon mixing monoclonal biotinylated antibody, the enzyme-labeled antibody and a serum containing the native antigen, reaction results between the native antigen and the antibodies, without competition or steric hindrance, to form a soluble sandwich complex.
    Analytical Method
    Quantitative
    产品特性
    The Quantitative Determination of Alpha-Fetoprotein (AFP) Concentration in Human Serum by a Microplate Chemiluminescence assay
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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.
    样本量
    25 μL
    板类型
    Pre-coated
    实验流程

    Specimien Collection and Preparation:

    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 redtop venipuncture tube without additives or anti-coagulants. Allow the blood to clot. 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, 0.050ml of the specimen is required.

    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. _x000E_

    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, con_x001F_trol or specimen into the assigned well. 3. Add 0.100 ml (100l) of the AFP Tracer Reagent to each well. It is very important to dispense all reagents close to the bottom of the coated well. 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, tap and 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. 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. Incubate for five (5) minutes in the dark. 10. Read the relative light units in each well for 0.2 1.0 seconds. The results should be read within thirty (30) minutes of adding the substrate solution.
    限制
    仅限研究用
  • 抗原 See all alpha Fetoprotein (AFP) CLIA Kits
    alpha Fetoprotein (AFP) (alpha-Fetoprotein (AFP))
    Abstract
    AFP 产品
    别名
    FETA CLIA Kit, HPAFP CLIA Kit, alpha fetoprotein CLIA Kit, alpha-fetoprotein CLIA Kit, AFP CLIA Kit, Afp CLIA Kit
    背景
    Alpha-Fetoprotein (AFP) is a glycoprotein with a molecular weight of 70 kDA. AFP shares considerable sequential homology with albumin and is normally produced during fetal development by the hepatocytes, yolk sac and to a lesser extent by the gastrointestinal tract. AFP appears as a major serum protein in the fetus, but its concentration declines rapidly towards birth. Serum concentrations reach a peak level of up to 10 mg/ml at twelve weeks of gestation (1). This peak level gradually decreases to less than 25 ng/ml after one year of postpartum. Thereafter, the levels reduce further to less than 10 ng/ml. Ever since its first reported association with tumors, by Tatarinov in 1964 based on his work with liver carcinomas, AFP has been a subject of discussion with relation to many different tumors. Abnormal AFP levels have been associated with hepatocellular carcinoma, ovarian cancer, gastrointestinal cancer and pulmonary cancer and, most recently, with non-seminomatous testicular cancer. Elevated levels of AFP are found in patients with primary heptatoma and yolk sac-derived germ tumors. AFP is the most useful marker for the diagnosis and management of hepatocellular carcinoma (2). AFP is also elevated in pregnant women. Presence of abnormally high AFP concentrations in pregnant women provides a risk marker for Down syndrome (3), In this method, AFP calibrator, patient specimen or control is first added to a streptavidin coated well. Biotinylated monoclonal and enzyme labeled antibodies (directed against distinct and different epitopes of AFP) are added and the reactants mixed. Reaction between the various AFP antibodies and native AFP forms a sandwich complex that binds with the streptavidin coated to the well. After the completion of the required incubation period, the enzyme-AFP antibody bound conjugate is separated from the unbound enzyme-AFP 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 measurable with a luminometer. The employment of several serum references of known alpha-fetoprotein (AFP) levels permits the 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 AFP concentration. _x000E_
    途径
    C21-Steroid Hormone Metabolic Process
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