serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Assay Length:
3.5h
Research Area:
Cytokine;Hematology;
Uniprot ID:
P20826
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 Mouse SCF. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Mouse SCF. 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 Mouse SCF, 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 Mouse SCF 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 (pg/mL)
OD
Corrected OD
1000.00
2.121
2.014
500.00
1.643
1.536
250.00
1.243
1.136
125.00
0.872
0.765
62.50
0.532
0.425
31.25
0.252
0.145
15.63
0.234
0.127
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 SCF and the recovery rates were calculated by comparing the measured value to the expected amount of SCF in samples.
Matrix
Recovery range
Average
serum(n=5)
96-104%
100%
EDTA plasma(n=5)
98-104%
102%
Heparin plasma(n=5)
95-107%
101%
線(xiàn)性
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of SCF and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.