应用备注 |
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说明 |
Information on standard material: |
样本量 | 100 μL |
实验时间 | 3 h |
板类型 | Pre-coated |
实验流程 | The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Hemopexin (HPX). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Hemopexin (HPX). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Hemopexin (HPX), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Hemopexin (HPX) in the samples is then determined by comparing the O.D. of the samples to the standard curve. |
试剂准备 |
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样品收集 |
Serum: Allow samples to clot for two hours at room temperature or overnight at 4°C before centrifugation for 20 minutes at approximately 1000 × g. Assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles. Plasma: Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000 × g within 30 minutes of collection. Remove plasma and assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles. Biological Fluids: Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at -20 °C or -80 °C for later use. Avoid repeated freeze/thaw cycles. |
样品制备 |
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实验流程 |
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结果分析 |
Average the duplicate readings for each standard, control, and samples and subtract the average zero standard optical density. Create a standard curve on log-log graph paper, with HPX concentration on the y-axis and absorbance on the x-axis. Draw the best fit straight line through the standard points and it can be determined by regression analysis. Using some plot software, for instance, curve expert 1.30, is also recommended. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor. |
实验精密度 |
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Hemopexin (HPX) were tested 20 times on one plate, respectively. |
限制 | 仅限研究用 |
注意事项 | The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material. |
注意事项 |
The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5 % within the expiration date under appropriate storage condition. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end. |
储存条件 | 4 °C |
储存方法 |
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有效期 | 6 months |
厂商提供的图像 |
有引用在: |
Aung, Dixon, Smith, Sweeney, Robson, Berney, Buxton, Green, Cook: "Novel regulatory roles of cAMP receptor proteins in fast-growing environmental mycobacteria." in: Microbiology (Reading, England), Vol. 161, Issue Pt 3, pp. 648-61, 2015 (PubMed).
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