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CA 19-9 ELISA 试剂盒

CA 19-9 适用: 人 Colorimetric Sandwich ELISA
产品编号 ABIN649061
发货至: 中国
  • 抗原 See all CA 19-9 ELISA试剂盒
    CA 19-9 (Gastrointestinal Cancer Antigen CA19-9 (CA 19-9))
    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 anti-CA19-9 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. 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 free 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. Human Serum References (1. 0 ml/vial). Six vials of human serum based reference calibrators at concentrations of 0 (A), 10 (B), 50 (C), 100 (D), 250 (E) and 500 (F) U/ml. Store at 2-8°C. A preservative has been added. Note: The standards, human serum based, were made using a greater than99% pure affinity purified preparation of CA 19-9. The preparation was calibrated against Centocor CA 19-9 IRMA test. B. CA19-9 Biotin Reagent (13 ml/vial). One vial of Anti-Human CA19-9 (MoAb)-Biotin reagent in a protein-stabilized matrix. A preservative has been added. Store at 2-8°C. C. CA19-9 Enzyme Reagent (13 ml/vial). One vial of Anti-Human CA19-9-HRP conjugate in a protein-stabilized matrix. A preservative has been added. Store at 2-8°C. D. Streptavidin Plate (96 wells). One 96-well microplate coated with streptavidin and packaged in an aluminum bag with a drying agent. Store at 2-8°C. E. Wash Solution Concentrate (20ml). One vial containing a surfactant in buffered saline. A preservative has been added. Store at 2-30°C. F. Substrate A (7. 0ml/vial). One bottle containing tetramethylbenzidine (TMB) in acetate buffer. Store at 2-8°C. G. Substrate B (7. 0ml/vial). One bottle containing hydrogen peroxide (H2O2) in acetate buffer. Store at 2-8°C. H. Stop Solution (8. 0ml/vial). One bottle containing a strong acid (1N HCl). Store at 2-8°C. 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. Note 3: Above reagents are for a single 96-well microplate.
    试剂未包括
    1. Pipette capable of delivering 25 & 50µl 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. Microplate washers or a squeeze bottle (optional). 4. Microplate Reader with 450nm and 620nm wavelength absorbance capability. 5. Absorbent Paper for blotting the microplate wells. 6. Plastic wrap or microplate cover for incubation steps. 7. Vacuum aspirator (optional) for wash steps. 9. Timer. 10. Quality control materials.
    Top Product
    Discover our top product CA 19-9 ELISA Kit
  • 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 licensed reagents. 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
    试剂准备
    1. Wash Buffer: Dilute contents of wash solution 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 vial labeled Solution A into the vial labeled Solution B. Place the yellow cap on the mixed reagent for easy identification. Mix and label accordingly. Store at 2-8°C. Note: Do not use the working substrate if it 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 redtop venipuncture tube without additives or anti-coagulants. Allow the blood to clot for samples. 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. 050ml of the specimen is required.
    结果分析

    A dose response curve is used to ascertain the concentration of CA19-9 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 CA19-9 concentration in U/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 CA19-9 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 U/ml) from the horizontal axis of the graph (the duplicates of the unknown may be averaged as indicated).

    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 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 (25 µl) of the appropriate serum reference, control or specimen into the assigned well. 3. Add 0. 100 ml (100µl) of the biotinylated labeled antibody 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 60 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 300µl 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 employed, fill each well by depressing the container (avoiding air bubbles) to dispense the wash. Decant the wash and Repeat two additional times. 8. Add 0. 100 ml (100µl) of the Ca19-9 Enzyme Reagent labeled antibody to each well. DO NOT SHAKE THE PLATE AFTER ENZYME ADDITION. 9. Cover and incubate 60 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 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 employed, fill each well by depressing the container (avoiding air bubbles) to dispense the wash. Decant the wash and Repeat two additional times. 12. Add 0. 100 ml (100µl) of working substrate solution to all wells (see Reagent Preparation Section). Always add reagents in the same order to minimize reaction time. DO NOT SHAKE THE PLATE AFTER SUBSTRATE ADDITION. 13. Incubate at room temperature for 15 minutes. 14. Add 0. 050ml (50µl) of stop solution to each well and gently mix for 15-20 seconds. 15. 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.
    储存条件
    4 °C/-20 °C
  • 抗原 See all CA 19-9 ELISA试剂盒
    CA 19-9 (Gastrointestinal Cancer Antigen CA19-9 (CA 19-9))
    别名
    CA 19-9 (CA 19-9 产品)
    背景
    Summary and Explanation of the test: A mucin type Sialyl Lewis Antigens group of glycoproteins (SLA) such as CA 19-9, 19-5 have been recognized as circulating cancer associated antigens for gastrointestinal cancer. The discovery of a monoclonal antibody clone (1116NS 19-9), which exhibited selective reactivity with human gastrointestinal carcinomas through the recognition of a carbohydrate determinant (CA 19-9) defined as a sialyl lacto-N-flucopenrose II, resulted in the successful purification and thus, determination of human gastrointestinal tumor associated glycoprotein antigen expressing CA 19-9 from colorectal carcinoma cell lines. Recently reports indicate that serum CA 19-9 level is frequently elevated in the circulation of patients with various gastrointestinal malignancies, such as pancreatic, colorectal, gastric and hepatic carcinomas. Together with CEA elevated CA 19-9 is suggestive of gallbladder disease. The tumor associated antigen may also be associated in some malignant conditions. Research studies demonstrate that serum CA 19-9 values may have utility in monitoring subjects with the above mentioned diagnosed malignancies. In this method, CA19-9 calibrator, patient specimen or control is first added to a streptavidin coated well. Biotinylated monoclonal antibody (specific for CA19-9) is added and the reactants mixed. Reaction between the CA19-9 antibodies and native CA19-9 forms complex that binds with the streptavidin coated to the well. The excess serum proteins are washed away via a wash step. Another enzyme labeled monoclonal antibody specific to CA19-9 is added to the wells. The enzyme labeled antibody binds to the CA19-9 already immobilized on the well through its binding with the biotinylated monoclonal antibody. Excess enzyme is washed off via a wash step. A color is generated by the addition of a substrate. The intensity of the color generation is directly proportional to the concentration of the CA19-9 in the sample. Intended Use: The Quantitative Determination of Cancer Antigen (CA 19-9) Concentration in Human Serum by a Microplate Immunoenzymometric assay. Q. C. Parameters: In order for the assay results to be considered valid the following criteria should be met: 1. The absorbance (OD) of calibrator F should be greater than 1.3. 2. Four out of six quality control pools should be within the established ranges.
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