This kit is based on Double antibody-Sandwich ELISA detection method and takes 4h assay time. The microplate provided in this kit has been precoated with anti F1+2 antibody. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add biotinylated detection antibody. Then, it binds with F1+2 bound to precoated antibody. Wash unbound components and add HRP-Streptavidin Conjugate (SABC). Wash unbound components again and add TMB substrate solution. Then, TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution. Read the O.D. absorbance at 450nm in a microplate reader. Calculate the concentration of F1+2 in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.
| Product Details | |
| Product Name | Human F1+2(Prothrombin Fragment 1+2) ELISA Kit |
| Alias | F1+2 ELISA Kit, Prothrombin Fragment 1+2 ELISA Kit |
| UniProt ID | P00734 |
| Sample Type | Serum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Other liquid samples |
| Detection Method | Sandwich ELISA, Double Antibody |
| Detection Wavelength | OD450 |
| Reaction Duration | 4 hours |
| Storage | 2-8°C(Sealed), Don't cryopreserve. |
| Specificity | Specifically binds with F1+2 , no obvious cross reaction with other analogues. |
Standard Curve
This product is detected by QC department and meets performance required in the manual. (Laboratory Humidity: 20%-60%; Temperature: 18°C -25°C; Equilibrate TMB substrate to 37°C before staining. After adding into the ELISA wells, incubate for 15min at 37°C in dark.)
Due to different assay environments and operations, assay data below and standard curve are provided for reference. Experimenters should establish standard curve according to their own assay.
| STD.(pg/ml) | OD-1 | OD-2 | Average |
| 0 | 0.077 | 0.079 | 0.078 |
| 46.875 | 0.14 | 0.144 | 0.142 |
| 93.75 | 0.194 | 0.2 | 0.197 |
| 187.5 | 0.311 | 0.321 | 0.316 |
| 375 | 0.534 | 0.55 | 0.542 |
| 750 | 0.916 | 0.942 | 0.929 |
| 1500 | 1.473 | 1.515 | 1.494 |
| 3000 | 2.12 | 2.182 | 2.151 |
Recovery
Add a certain amount of F1+2 into the sample. Calculate the recovery by comparing the measured value with the expected amount of F1+2 in the sample
| Sample Type | Recovery Range(%) | Average(%) |
| serum(n=10) | 86-100 | 91 |
| EDTA plasma(n=10) | 89-100 | 93 |
| Heparin plasma(n=10) | 90-104 | 97 |
Linearity
Dilute the sample with a certain amount of F1+2 at 1:2, 1:4 and 1:8 to get the recovery range.
| Sample Type | 1:2 | 1:4 | 1:8 |
| serum(n=10) | 93-99% | 88-99% | 88-100% |
| EDTA plasma(n=10) | 86-96% | 84-95% | 83-97% |
| Heparin plasma(n=10) | 86-96% | 86-98% | 88-99% |
Precision(%)
Intra-assay Precision: samples with low, medium and high concentration are tested 20 times on the same plate.
Inter-assay Precision: samples with low, medium and high concentration are tested 20 times on three different plates.
| Item | Intra-assay Precision | Inter-assay Precision | ||||
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/ml) | 91.8 | 369.1 | 1464 | 93.48 | 374.6 | 1489 |
| Standard deviation | 4.36 | 18.45 | 73.93 | 4.64 | 18.66 | 74.15 |
| CV(%) | 4.75 | 5 | 5.05 | 4.96 | 4.98 | 4.98 |
Stability
Perform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data.
| ELISA kit(n=5) | 37°C for 1 month | 2-8°C for 6 months |
| Average(%) | 80 | 95-100 |
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