Progesterone ELISA 试剂盒
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- 抗原 See all Progesterone ELISA试剂盒
- Progesterone
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适用
- 人
- 检测方法
- Colorimetric
- 实验类型
- Competition ELISA
- 应用范围
- ELISA
- 原理
- Competitive Enzyme Immunoassay (TYPE 7): The essential reagents required for a enzyme immunoassay include antibody, enzyme-antigen conjugate and native antigen. Upon mixing biotinylated antibody, enzyme-antigen conjugate and a serum containing the native antigen, a competition reaction results between the native antigen and the enzyme-antigen conjugate for a limited number of antibody binding sites. The enzyme activity in the antibody bound fraction is inversely proportional to the native 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. Progesterone Calibrators (1ml/vial). Seven vials of serum reference for progesterone at concentrations of 0 (A), 0. 3 (B), 2. 0 (C), 5. 0 (D), 15 (E), 30 (F) and 60. 0 (G) ng/ml. Store at 2-8°C. A preservative has been added. The calibrators can be expressed in molar concentrations (nM/L) by multiplying by 3. 18. For example: 1ng/ml x 3. 18 equal 3. 18 nM/LB. Progesterone Enzyme Reagent (6. 0 ml/vial). One vial of ProgesterOne (Analog)-horseradish peroxides (HRP) conjugate in a protein stabilizing matrix with red dye. Store at 2-8°C. C. Progesterone Biotin Reagent (6. 0 ml). One bottle of reagent contains anti-Progesterone biotinylated purified rabbit IgG conjugate in buffer, yellow dye and preservative. Store at 2-8°C. D. Streptavidin Coated Plate (96 wells). One 96-well microplate coated with 1. 0 µg/ml streptavidin and packaged in an aluminum bag with a drying agent. Store at 2-8°C. E. Wash Solution Concentrate (20ml). One vial contains a surfactant in buffered saline. A preservative has been added. Store at 2-30°C. F. Substrate Reagent (12ml/vial). One bottle contains tetramethylbenzidine (TMB) and hydrogen peroxide (H2O2) in buffer. Store at 2-8°C. G. Stop Solution (8ml/vial). One vial contains a strong acid (H2SO4). Store at 2-30°C. H. 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 25ml and 50ml 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. Adjustable volume (200-1000µl) Dispenser(s) for conjugate. 4. Microplate washer or a squeeze bottle (optional). 5. Microplate Reader with 450nm and 620nm wavelength absorbance capability. 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. 10. Quality control materials.
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Progesterone ELISA Kit
适用: Various Species Colorimetric Competition ELISA 1.23 ng/mL - 100 ng/mL Plasma, Serum
Progesterone ELISA Kit适用: Various Species Colorimetric Competition ELISA 0.31 ng/mL - 20 ng/mL Plasma, Saliva, Serum, Urine
Progesterone ELISA Kit适用: 小鼠 Colorimetric Competition ELISA 0.313 ng/mL - 20 ng/mL Plasma, Serum, Tissue Homogenate
Progesterone ELISA Kit适用: Various Species Colorimetric Sandwich ELISA Fecal, Tissue Culture Medium, Urine
Progesterone ELISA Kit适用: 大鼠 Colorimetric Competition ELISA 0.313 ng/mL - 20 ng/mL Plasma, Serum, Tissue Homogenate
Progesterone ELISA Kit适用: Various Species Colorimetric Competition ELISA 0.313 ng/mL - 20 ng/mL Plasma, Serum, Tissue Homogenate
Progesterone ELISA Kit适用: 小鸡 Colorimetric Competition ELISA 0.313 ng/mL - 20 ng/mL Plasma, Serum, Tissue Homogenate
Progesterone ELISA Kit适用: 绵羊 Colorimetric Competition ELISA 0.313 ng/mL - 20 ng/mL Plasma, Serum, Tissue Homogenate
Progesterone ELISA Kit适用: Camel Colorimetric Competition ELISA 0.313 ng/mL - 20 ng/mL Plasma, Serum, Tissue Homogenate
Progesterone ELISA Kit适用: Pig Colorimetric Competition ELISA 0.313 ng/mL - 20 ng/mL Plasma, Serum, Tissue Homogenate
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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 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
- 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. Diluted buffer can be stored at room temperature (20-27°C) for up to 60 days. Note: Do not use the working substrate if it looks blue.
- 样品收集
- The specimens shall be blood, serum or heparanised plasma in type and taken with the usual precautions in the collection of venipuncture samples. For accurate comparison to establish normal values, a fasting morning serum sample should be obtained. The blood should be collected in a redtop (with or without gel additives) venipuncture tube or for plasma use evacuated tube(s) containing heparin. Allow the blood to clot for serum samples. Centrifuge the specimen to separate the serum or plasma 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 progesterone 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 progesterone concentration in ng/ml on linear graph paper (do not average the duplicates of the serum references before plotting). 3. Connect the points with a best-fit curve. 4. To determine the concentration of progesterone 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).
- 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 (25 µl) of the appropriate serum reference, control or specimen into the assigned well. 3. Add 0. 050 ml (50µl) of Progesterone Enzyme Reagent to all wells. 4. Swirl the microplate gently for 10-20 seconds to mix. 5. Add 0. 050 ml (50µl) of working Progesterone Biotin Reagent to all wells. 6. Swirl the microplate gently for 10-20 seconds to mix. 7. Cover and incubate for 60 minutes at room temperature. 8. Discard the contents of the microplate by decantation or aspiration. If decanting, blot the plate dry with absorbent paper. 9. 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. 10. Add 0. 100 ml (100µl) of Substrate solution to all wells. Always add reagents in the same order to minimize reaction time differences between wells. DO NOT SHAKE THE PLATE AFTER SUBSTRATE ADDITION. 11. Incubate at room temperature for 20 minutes. 12. Add 0. 050ml (50µl) of stop solution to each well and gently mix for 15-20 seconds. Always add reagents in the same order to minimize reaction time differences between wells. 13. Read the absorbance in each well at 450nm (using a reference wavelength of 620-630nm. The results should be read within 30 minutes of adding the stop solution. Note: Dilute the samples suspected of concentrations higher than 60ng/ml 1:5 and 1:10 with progesterone 0 ng/ml calibrator or male patient serum pools with a known low value for progesterone.
- 储存条件
- 4 °C/-20 °C
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- 抗原 See all Progesterone ELISA试剂盒
- Progesterone
- Abstract
- Progesterone 产品
- 物质类
- Hormone
- 背景
- Summary and Explanation of the test: Measurement of progesterone in serum or plasma is considered to be the most reliable way to assess its rate of production. Progesterone is a steroid hormone, which plays an important role in the preparation for and maintenance of pregnancy. It is synthesized from cholesterol via pregnenolone, then rapidly metabolized to pregnanediol primarily in the liver. The ovary and placenta are the major production sites, but a small amount is also produced by the adrenal cortex in both men and women. Circulating progesterone levels, which are characteristically low during the follicular phase, increase sharply during the luteal phase of menstrual cycles, reaching a maximum approximately 5 to 10 days after the midcycle LH peak. 12 Unless pregnancy occurs, a steep decline to follicular levels sets in about 4 days before the next menstrual period. This pattern constitutes the rationale behind the well established use of serum progesterone measurements as a simple and reliable method for ovulation detection. For routine measurements, immunoassays using steroid specific antibodies are preferred. Initial immunoassays, for serum progesterone, used organic solvents to remove the steroid from endogenous binding proteins such as corticosteroid binding globulin (CBG) and albumin. Direct measurement of progesterone in serum or plasma is considered to be the method of choice for routine applications. Both RIA and EIA (and some FIA) are available in the market. Since RIA involves handling radioactivity and causes radioactive waste disposal issues, various non-isotopic methods have replaced the RIA. These methods use very specific antibodies to determine levels of progesterone in circulation. Progesterone ELISA kit use a specific anti-progesterone antibody, and do not require sample extraction of serum or plasma. Cross-reactivity to other naturally occurring and structurally related steroids is low. The employment of several serum references of known progesterone concentration permits construction of a graph of activity and concentration. From comparison to the dose response curve, an unknown specimen's activity can be correlated with progesterone concentration. Intended Use: The Quantitative Determination of Progesterone Concentration in Human Serum or Plasma by a Microplate Enzyme Immunoassay. 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 0 ng/ml should be greater than 1.3. 2. Four out of six quality control pools should be within the established ranges.
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