CEA ELISA 试剂盒
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- 抗原 See all CEA ELISA试剂盒
- CEA (Carcinoembryonic Antigen Gene Family (CEA))
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适用
- 人
- 检测方法
- Colorimetric
- 实验类型
- Sandwich ELISA
- 应用范围
- ELISA
- 原理
- Immunoenzymometric assay (TYPE 3): 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-CEA 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. TAfter equilibrium is attained, the antibodybound fraction is separated from unbound antigen by decantation or aspiration. The enzyme activity in the antibodybound fraction is directly proportional to the native antigen concentration. By utilizing several different serum references of known antigen values, a dose response curve can be generated from which the antigen concentration of an unknown can be ascertained.
- Analytical Method
- Quantitative
- 组件
- A. Carcinoembryonic antigen (CEA) (1ml/vial). Six vials of references CEA Antigen at levels of 0 (A), 5 (B), 10 (C), 25 (D), 50 (E) and 250 (F) ng/ml. Store at 2-8°C. A preservative has been added. Note: The standards, human serum based, were calibrated using a reference preparation, which was assayed against the 1st International Reference Preparation (IRPNumber 73/601). B. CEA Enzyme Reagent (13ml/vial): One vial containing enzyme labeled antibody, biotinylated monoclonal mouse IgG in buffer, dye, and preservative. Store at 2-8°C. C. Streptavidin Coated 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. D. Wash Solution Concentrate (20 ml). One vial containing a surfactant in buffered saline. A preservative has been added. Store at 2-30°C. E. Substrate A (7ml/vial). One bottle containing tetramethylbenzidine (TMB) in buffer. Store at 2-8°C. F. Substrate B (7ml/vial). One bottle containing hydrogen peroxide (H2O2) in buffer. Store at 2-8°C. G. Stop Solution (8ml/vial). One bottle containing a strong acid (1N HCl). Store at 2-30°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. Note 3: Above reagents are for a single 96-well microplate.
- 试剂未包括
- 1. Pipette (s) capable of delivering 25µl,& 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. 8. Timer. 9. Quality control materials.
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Carcinoembryonic Antigen Gene Family (CEA) ELISA Kit
CEA 适用: 小鼠 Colorimetric Sandwich ELISA 31.2 pg/mL - 2000 pg/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
Carcinoembryonic Antigen Gene Family (CEA) ELISA KitCEA 适用: 人 Colorimetric Sandwich ELISA 19.6 pg/mL - 1250 pg/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
Carcinoembryonic Antigen Gene Family (CEA) ELISA KitCEA 适用: 大鼠 Colorimetric Sandwich ELISA 0.312 ng/mL - 20 ng/mL Plasma, Serum, Tissue Homogenate
Carcinoembryonic Antigen Gene Family (CEA) ELISA KitCEA 适用: 大鼠 Colorimetric Sandwich ELISA 0.16 ng/mL - 10 ng/mL Cell Culture Supernatant, Plasma, Serum
Carcinoembryonic Antigen Gene Family (CEA) ELISA KitCEA 适用: 大鼠 Colorimetric Sandwich ELISA 0.312 ng/mL - 20 ng/mL Cell Lysate, Plasma, Serum, Tissue Homogenate
Carcinoembryonic Antigen Gene Family (CEA) ELISA KitCEA 适用: 人 Colorimetric Sandwich ELISA 312 pg/mL - 20000 pg/mL Cell Culture Supernatant, Plasma (EDTA - heparin), Serum
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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 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
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- 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 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 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. 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
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A dose response curve is used to ascertain the concentration of Carcinoembryonic antigen 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 CEA concentration in ng/ml on linear graph paper (do not average the duplicates of the serum references before plotting). 3. Draw the best-fit curve through the plotted points. 4. To determine the concentration of CEA 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 assays may also be used for the data reduction.
- 限制
- 仅限研究用
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- 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 (25µl) of the appropriate serum reference, control or specimen into the assigned well. 3. Add 0. 100 ml (100µl) of the CEA Enzyme 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 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 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. 8. 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 differences between wells. DO NOT SHAKE THE PLATE AFTER SUBSTRATE ADDITION. 9. Incubate at room temperature for 15 minutes. 10. 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. 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
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- 抗原 See all CEA ELISA试剂盒
- CEA (Carcinoembryonic Antigen Gene Family (CEA))
- 别名
- Carcinoembryonic Antigen (CEA) (CEA 产品)
- 别名
- RGD1311281 ELISA Kit, carcinoembryonic antigen-related cell adhesion molecule 20 ELISA Kit, carcinoembryonic antigen gene family ELISA Kit, Ceacam20 ELISA Kit, Cea ELISA Kit
- 背景
- Summary and Explanation of the test: Carcinoembryonic antigen (CEA) is a glycoprotein with a molecular weight of 180 kDA. CEA is the first of the so-called carcinoembryonic proteins that was discovered in 1965 by Gold and Freeman (1). CEA is the most widely used marker for gastrointestinal cancer. Although CEA is primarily associated with colorectal cancers, other malignancies that can cause elevated levels of CEA include breast, lung, stomach, pancreas, ovary and other organs. Benign conditions that cause significantly higher than normal levels include inflammation of lung and gastrointestinal (GI) tract and benign liver cancer (2,3). Heavy Smokers, as a group, have higher than normal baseline concentration of CEA. In this method, CEA 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 CEA) are added and the reactants mixed. Reaction between the various CEA antibodies and native CEA forms a sandwich complex that binds with the streptavidin coated to the well. After the completion of the required incubation period, the enzyme-CEA antibody bound conjugate is separated from the unbound enzyme-CEA 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 color. The employment of several serum references of known carcinoembryonic antigen (CEA) 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 CEA concentration. Intended Use: The Quantitative Determination of Carcinoembryonic Antigen (CEA) 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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