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Human 5-NT(5-Nucleotidase) ELISA Kit
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Product No

FY-EH1551

Sample Type

Serum, Plasma, Tissue homogenate and Other biological samples

Assay Time

3.5H

Price($)

280 400

Size

48T 96T

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DataSheet DataSheet MSDS MSDS
Product Details
Product Name in English:
Human 5-NT(5-Nucleotidase) ELISA Kit
Species:
Human
Alternative Name:
NT5E, CD73, E5NT, NT, NT5, NTE, eN, eNT, ecto-5'-nucleotidase
Uniport ID:
P21589
Assay Type:
Sandwich
Standard:
20ng/mL
Sensitivity:
0.19 ng/mL
Detection Range:
0.32-20ng/mL
Assay Time:
3.5H
Sample Type:
Serum, Plasma, Tissue homogenate and Other biological samples
Research:
Signal transduction;Tumor immunity;Infection immunity;
Test Principle:
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 5-NT. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to 5-NT. 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 5-NT, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of 5-NT in the samples is then determined by comparing the OD of the samples to the standard curve.
Technical Data
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
Concentration OD1 OD2 Average OD Corrected
20 2.307 2.31 2.309 2.284
10 1.513 1.528 1.521 1.49
5 0.882 0.89 0.886 0.859
2.5 0.399 0.402 0.401 0.376
1.25 0.22 0.221 0.221 0.197
0.63 0.12 0.121 0.121 0.097
0.32 0.073 0.073 0.073 0.05
0 0.023 0.023 0.023 0
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level were tested on 3 different plates, 20 replicates in each plate, respectively.
Item Intra-assay Precision Intra-assay Precision
Sample 3 3
Replicate 9 18
CV(%) 5 8
Rate of recovery
The recovery of spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
Sample Type Range(%) Average Recovery(%)
Serum (n=8) 80-94 87
EDTA plasma (n=8) 95-107 101
Cell culture media (n=8) 82-95 88
Linear
Samples were spiked with high concentrations of target proteins and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
1:2 Range (%) 88-102 90-99 82-97
Average (%) 93 97 86
1:4 Range (%) 93-105 90-99 88-103
Average (%) 101 91 103
1:8 Range (%) 91-99 95-106 83-94
Average (%) 98 106 84
1:16 Range (%) 83-96 87-101 92-105
Average (%) 91 100 100
Assay Procedures
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