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Luteinizing Hormone CLIA Kit

LH 适用: 人 Chemiluminescent Sandwich ELISA
产品编号 ABIN504782
发货至: 中国
  • 抗原 See all Luteinizing Hormone (LH) CLIA Kits
    Luteinizing Hormone (LH)
    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-LH 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 Luteinizing Hormone Concentration in Human Serum by a Microplate Chemiluminescence Assay (CLIA)
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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
    50 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 red-top 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, 0.100ml 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 microplate 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. Add 0.100 ml (100l) of LH-Enzyme Conjugate solution to all wells. 4. Swirl the plate 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. 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 at room temperature in the dark for five (5) min 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 substrate solution.
    限制
    仅限研究用
  • 抗原 See all Luteinizing Hormone (LH) CLIA Kits
    Luteinizing Hormone (LH)
    Abstract
    LH 产品
    别名
    Plod CLIA Kit, procollagen-lysine, 2-oxoglutarate 5-dioxygenase 1 CLIA Kit, Plod1 CLIA Kit
    背景
    Luteinizing hormone (LH) is a glycoprotein consisting of two subunits with a molecular mass of 30,000 daltons. The (-subunit is similar to other pituitary hormones (follicle stimulating hormone (FSH), thyroid stimulating hormone (TSH) and chorionic gonadotropin (CG)( while the (-subunit is unique. The (-subunit confers the biological activity to the molecule. The (-subunit consists of 89 amino acid residues while the (-subunit contains 129 amino acids. The carbohydrate content is between 15% and 30%. The clinical usefulness of the measurement of luteinizing hormone (LH) in ascertaining the homeostasis of fertility regulation via the hypothalamic - pituitary - gonadal axis has been well established (1,2). In addition, the advent of in vitro fertilization (IVF) technology to overcome infertility-associated problems has provided the impetus for rapid improvement in LH assay methodology from the technically demanding bioassay (3) to the procedurally simple and rapid immunoenzymometric assays. In this method, LH 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 LH) are added and the reactants mixed. Reaction between the various LH antibodies and native LH forms a sandwich complex that binds with the streptavidin coated to the well. After the completion of the required incubation period, the enzyme-luteinizing hormone antibody bound conjugate is separated from the unbound enzyme-luteinizing hormone 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 luteinizing hormone levels permits 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 luteinizing hormone concentration.
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