Description
Product Name
Human TNF-α(Tumor Necrosis Factor Alpha) ELISA Kit
Species
Human
Alternative Names
DIF, TNF-alpha, TNFA, TNFSF2, TNF-A, TNFα
Detection method
0.47 pg/mL
Standard
50pg/mL
Detection Range
0.78-50pg/mL
Sample type
Serum, Plasma and Other biological fluids,Sample Volume= 100μL
Reaction time
3.5H
Research Area
Cancer, Immunology
Test principle
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 TNF-α. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human TNF-α and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human TNF-α, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. 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 nm ± 2 nm. The OD value is proportional to the concentration of Human TNF-α. You can calculate the concentration of Human TNF-α in the samples by comparing the OD of the samples to the standard curve.
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.
(pg/mL)
OD
Corrected
50.00
2.313
2.284
25.00
1.519
1.490
12.50
0.888
0.859
6.25
0.405
0.376
3.13
0.226
0.197
1.56
0.126
0.097
0.78
0.079
0.050
0.00
0.029
0.000
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.
Intra-assay Precision
Inter-assay Precision
Sample
1
2
3
1
2
3
n
20
20
20
20
20
20
Mean(pg/mL)
8.75
10.16
26.04
11.06
26.33
53.20
Standard deviation
0.69
4.94
11.77
0.50
4.57
| 11.83 |
C V (%)
5.48
7.36
5.57
7.17
4.01
5.62
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)
80-90
85
EDTA plasma (n=8)
84-106
100
Cell culture media (n=8)
87-93
90
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 (%)
85-94
93-102
80-93
Average (%)
90
94
92
1:4
Range (%)
87-96
93-102
88-97
Average (%)
91
94
92
1:8
Range (%)
82-95
80-92
86-102
Average (%)
92
90
93
1:16
Range (%)
81-93
87-98
85-99
Average (%)
90
92
93
Reviews
There are no reviews yet.