Catalog No: RE2358HF
MF-Human TNF-β(Tumor Necrosis Factor Beta) ELISA Kit
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48T

$299

96T

$399

Product Details

Species:

Human

Alternative Name:

LT, LTA, TNFB, TNFSF1, TNF Superfamily,Member 1, Lymphotoxin Alpha, TNFβ

Uniport ID:

P01374

Assay Type:

Sandwich

unit:

pg/mL

Standard:

2000pg/mL

Sensitivity:

18.75 pg/mL

Detection Range:

31.25-2000pg/mL

Assay Time:

3H

Sample Type:

Serum, Plasma and Tissue homogenate and Other biological samples;Sample volume:25μL

Research:

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-β. Samples (or Standards) are added to the micro ELISA plate wells and combined with the specific antibody and biotinylated detection antibody specific for Human TNF-β. Then 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-β, 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. The OD value is proportional to the concentration of Human TNF-β. You can calculate the concentration of Human TNF-β in the samples 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.
pg/mL OD Corrected
2000 2.097 2.01
1000 1.209 1.122
500 0.709 0.622
250 0.411 0.324
125 0.272 0.185
62.5 0.163 0.076
31.25 0.127 0.04
0 0.087 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.

Intra-assay Precision Inter-assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean(ng/mL) 29.54 194.06 447.97 31.75 186.01 394.67
Standard deviation 1.52 9.52 22.43 1.07 12.94 18.65
C V (%) 6.66 7.07 6.17 7.43 6.4 5.48

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) 82-95 88
EDTA plasma (n=8) 80-94 87
Cell culture media (n=8) 89-101 95

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 (%) 85-103 80-92 85-94
Average (%) 95 80 85
1:4 Range (%) 83-96 96-105 90-99
Average (%) 96 103 99
1:8 Range (%) 95-102 86-97 82-101
Average (%) 100 95 99
1:16 Range (%) 87-98 85-101 91-102
Average (%) 96 87 102

Assay Procedures

Description