serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Assay Length:
3.5h
Research Area:
Cytokine;Tumor immunity;
Uniprot ID:
P58294
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 Human EG-VEGF. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human EG-VEGF. 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 Human EG-VEGF, 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 Human EG-VEGF in the samples is then determined by comparing the OD of the samples to the standard curve.
標(biāo)準(zhǔn)曲線
Concentration (pg/mL)
OD
Corrected OD
1000.00
2.266
2.083
500.00
1.803
1.620
250.00
1.386
1.203
125.00
0.971
0.788
62.50
0.630
0.447
31.25
0.414
0.231
15.63
0.252
0.069
0.00
0.183
0.000
精密度
Intra-assay Precision (Precision within an assay):CV%<8%
Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-assay Precision (Precision between assays):CV%<10%
Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.
回收率
Matrices listed below were spiked with certain level of recombinant EG-VEGF and the recovery rates were calculated by comparing the measured value to the expected amount of EG-VEGF in samples.
Matrix
Recovery range
Average
serum(n=5)
82-95%
88%
EDTA plasma(n=5)
85-97%
91%
Heparin plasma(n=5)
79-93%
86%
線性
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of EG-VEGF and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.