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 IFABP/FABP2. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human IFABP/FABP2. 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 IFABP/FABP2, 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 IFABP/FABP2 in the samples is then determined by comparing the OD of the samples to the standard curve.
標(biāo)準(zhǔn)曲線(xiàn)
Concentration (ng/mL)
OD
Corrected OD
10.00
2.035
1.928
5.00
1.593
1.486
2.50
1.183
1.076
1.25
0.752
0.645
0.63
0.546
0.439
0.32
0.354
0.247
0.16
0.232
0.125
0.00
0.107
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 IFABP/FABP2 and the recovery rates were calculated by comparing the measured value to the expected amount of IFABP/FABP2 in samples.
Matrix
Recovery range
Average
serum(n=5)
82-95%
88%
EDTA plasma(n=5)
80-95%
87%
Heparin plasma(n=5)
86-99%
92%
線(xiàn)性
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of IFABP/FABP2 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.