|
Product Name |
Rat F1+2(Prothrombin Fragment 1+2) ELISA Kit |
|
Standard |
200 ng/mL |
|
Sample Type |
serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids |
|
Assay Length |
3.5h |
|
Research Area |
Hematology |
Standard Curve
|
Concentration (ng/mL) |
OD |
Corrected OD |
|
200.00 |
2.268 |
2.174 |
|
100.00 |
1.608 |
1.514 |
|
50.00 |
1.251 |
1.157 |
|
25.00 |
0.834 |
0.740 |
|
12.50 |
0.521 |
0.427 |
|
6.25 |
0.331 |
0.237 |
|
3.13 |
0.175 |
0.081 |
|
0.00 |
0.094 |
0.000 |
Precision
- 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.
Recovery
Matrices listed below were spiked with certain level of recombinant F1+2 and the recovery rates were calculated by comparing the measured value to the expected amount of F1+2 in samples.
|
Matrix |
Range(%) |
Average Recovery(%) |
|
Serum (n=8) |
87-95% |
91% |
|
EDTA plasma (n=8) |
93-107% |
100% |
|
Heparin plasma(n=5) |
87-99% |
93% |
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of F1+2 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
|
Matrix |
1:2 |
1:4 |
1:8 |
1:16 |
|
Serum (n=5) |
92-101% |
87-98% |
85-92% |
79-96% |
|
EDTA plasma(n=5) |
78-92% |
96-105% |
87-98% |
95-102% |
|
Heparin plasma(n=5) |
85-94% |
87-101% |
86-97% |
88-95% |
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 Rat F1+2. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rat F1+2. 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 Rat F1+2, 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 Rat F1+2 in the samples is then determined by comparing the OD of the samples to the standard curve.
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