产品名称 | FS-Human 5-NT(5-Nucleotidase) ELISA Kit | 反应种属 | Human |
Uniprot ID | P21589 | 别称 | 5' nucleotidase (CD73), 5'-NT, 5-NT, 5'-Nucleotidase, 5'-nucleotidase, ecto (CD73), CD73 antigen, CD73, E5NT, EC 3.1.3.5, Ecto-5'-nucleotidase, eN, eNT, NT, NT55'-nucleotidase, NT5E, NTE, Purine 5-Prime-Nucleotidase |
检测方法 | Sandwich | 灵敏度 | 2.86 pg/mL |
标准品 | 2000pg/mL | 检测范围 | 31.25-2000pg/mL |
样品类型 | Serum, Plasma, Tissue homogenate and Other biological samples;Sample Volume=50μL | ||
检测时间 | 1.5H | 研究领域 | Signal transduction;Tumor immunity;Infection immunity; |
测试原理 | This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human 5-NT. Samples (or Standards) and Horseradish Peroxidase (HRP) linked antibody specific for Human 5-NT are added to the micro ELISA plate wells. Human 5-NT in samples (or standards) combines with the coated antibody and HRP linked detection antibody special to Human 5-NT. Excess conjugate and unbound sample or standard are washed from the plate. The substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450±2 nm. The OD value is proportional to the concentration of Human 5-NT. The concentration of Human 5-NT in the samples is then determined by comparing the OD of the samples to the standard curve. |
由于实验操作条件的不同(如操作者、移液技术、洗板技术和稳定条件等),标准曲线的OD值会有所差异。以下数据和曲线仅供参考,实验者需根据自己的实验建立标准曲线。
(pg/mL) | OD | Corrected |
---|---|---|
2000.00 | 2.413 | 2.355 |
1000.00 | 1.415 | 1.357 |
500.00 | 0.850 | 0.792 |
250.00 | 0.494 | 0.436 |
125.00 | 0.291 | 0.233 |
62.50 | 0.179 | 0.121 |
31.25 | 0.112 | 0.054 |
0.00 | 0.058 | 0.000 |
板内精密度:低浓度样本,中浓度样本和高浓度样本分别在1块板子上检测20次。
板间精密度:低浓度样本,中浓度样本和高浓度样本分别在3块板子上检测20次。
Intra-assay Precision | Inter-assay Precision | |||||
---|---|---|---|---|---|---|
Sample | 1 | 2 | 3 | 1 | 2 | 3 |
n | 20 | 20 | 20 | 20 | 20 | 20 |
Mean(pg/mL) | 1723.734 | 977.440 | 889.308 | 450.374 | 444.014 | 373.350 |
Standard deviation | 153.730 | 90.770 | 32.417 | 9.370 | 11.193 | 10.157 |
C V (%) | 10.545 | 6.408 | 10.217 | 6.444 | 10.177 | 9.176 |
分别往5个不同样本中添加已知浓度的目标蛋白,做回收实验,得出回收率范围和平均回收率。
Sample Type | Range(%) | Average Recovery(%) |
---|---|---|
Serum (n=8) | 90-110 | 96 |
EDTA plasma (n=8) | 92-110 | 110 |
Cell culture media (n=8) | 92-110 | 107 |
分别往5个样本中添加已知浓度的目标蛋白,做回收实验,得出回收率范围及平均回收率。将5个样本分别稀释2倍,4倍,8倍,16倍做回收实验,得出回收率范围及平均回收率。
Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
---|---|---|---|---|
1:2 | Range (%) | 90-106 | 94-104 | 91-108 |
Average (%) | 102 | 99 | 93 | |
1:4 | Range (%) | 92-109 | 93-103 | 93-110 |
Average (%) | 96 | 105 | 108 | |
1:8 | Range (%) | 94-106 | 93-112 | 92-114 |
Average (%) | 103 | 92 | 94 | |
1:16 | Range (%) | 92-113 | 92-115 | 94-98 |
Average (%) | 111 | 98 | 97 |