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| Product Name | Mouse Coagulation Factor VII (FVII) ELISA Kit |
| Catalog No. | CFTK-HMM-0007 |
| Test Species | Mouse |
| Application | This kit is for the in vitro quantitative analysis of mouse coagulation factor VII (FVII) in serum, plasma, tissue homogenates and related liquid samples. |
| Shelf Life | 6 months |
| Storage | 2-8°C |
| Detection Principle | The kit uses a double-antibody one-step sandwich enzyme-linked immunosorbent assay (ELISA). To the coated microtiter wells pre-coated with mouse capture antibody for coagulation factor VII (FVII), the specimen, standard, and HRP-labeled detection antibody were added sequentially, incubated and washed thoroughly. The color was developed with the substrate TMB, which was converted to blue by catalysis of peroxidase and to final yellow by the action of acid. The color shade is positively correlated with mouse coagulation factor VII (FVII) in the sample. The absorbance (OD value) is measured at 450 nm with an enzyme meter to calculate the concentration of the sample. |
| Sample Processing | 1. Serum: Place whole blood specimens collected in serum separator tubes at room temperature for 2 hours or overnight at 4°C, then centrifuge at 1000×g for 20 minutes and remove the supernatant, or store the supernatant at -20°C or -80°C, but avoid repeated freezing and thawing. 2. Plasma: Collect the specimen with EDTA or heparin as anticoagulant and centrifuge the specimen at 1000×g for 15 minutes at 2-8°C within 30 minutes after collection, and then remove the supernatant for testing, or store the supernatant at -20°C or -80°C, but avoid repeated freezing and thawing. 3. Tissue Homogenization: Rinse the tissue with pre-cooled PBS (0.01M, pH=7.4) to remove residual blood (lysed erythrocytes in the homogenate will affect the measurement), weigh the tissue, and then cut the tissue into pieces. Mix the minced tissue with the corresponding volume of PBS (generally 1:9 weight-to-volume ratio, e.g., 1g of tissue sample corresponds to 9mL of PBS, the specific volume can be adjusted according to the experimental needs, and make a record. It is recommended to add protease inhibitors to PBS) into a glass homogenizer and grind it thoroughly on ice. In order to further lyse the tissue cells, the homogenate can be ultrasonically broken, or repeatedly frozen and thawed. Centrifuge the homogenate at 5000×g for 5-10 minutes and take the supernatant for testing. 4. Cell Culture Supernatant or Other Biological Specimens: Please centrifuge the supernatant at 1000×g for 20 minutes, and then take the supernatant for testing, or store the supernatant at -20°C or -80°C, but should avoid repeated freezing and thawing. Note: Hemolyzed specimens are not suitable for this test. |
| Self-contained Reagents / Instruments / Consumables | Enzyme labeler (450 nm) High-precision spikers and tips: 0.5-10 µL, 2-20 µL, 20-200 µL, 200-1000 µL 37°C thermostat Distilled or deionized water |
| Standard Concentration | 8, 4, 2, 1, 0.5, 0.25 IU/mL |
| Reagent Preparation | The kit should be equilibrated at room temperature before use when removed from the refrigerated environment. Dilution of 20 x Wash Buffer: distilled water is diluted 1:20, i.e. 1 part 20 x Wash Buffer to 19 parts distilled water. |
| Procedures | 1. Remove the required plates from the aluminum foil pouch after equilibrating at room temperature for 20 min, and seal the remaining plates in a self-sealing bag and return them to 4°C. 2. Set up standard wells and sample wells, add 50 µL of standards of different concentrations to each standard well. 3. Add 50 µL of the sample to be tested into the sample wells; do not add to the blank wells. 4. Except for the blank wells, add 100 µL of horseradish peroxidase (HRP)-labeled detection antibody to each of the standard and sample wells, seal the reaction wells with a sealing membrane, and incubate for 60 min at 37°C in a water bath or thermostat. 5. Discard the liquid, pat dry on absorbent paper, fill each well with washing solution (350 µL), let stand for 1 min, shake off the washing solution, pat dry on absorbent paper, and repeat the plate washing for 5 times (plate washer can also be used to wash the plate). 6. Add 50 µL of substrate A and B to each well and incubate at 37°C for 15 min. 7. Add 50 µL of termination solution to each well, and measure the OD value of each well at 450 nm within 15 min. |
| Calculation of Results | The experimental results were calculated by taking the OD value of the measured standard as the horizontal coordinate and the concentration value of the standard as the vertical coordinate, drawing the standard curve on the coordinate paper or using the relevant software and obtaining the linear regression equation, substituting the OD value of the samples into the equation and calculating the concentration of the samples. |
| Detection Range | 0.25 IU/mL-8 IU/mL |
| Sensitivity | < 0.1 IU/mL |
| Specificity | Does not cross-react with other soluble structural analogs. |
| Repeatability | The intraplate coefficient of variation is less than 10% and the interplate coefficient of variation is less than 15%. |
| Note | 1. After a large number of normal specimens, the normal concentration values of the specimens are within the detection range provided by the kit, and 50 µL of the sample can be sampled directly during the experiment. When the value of some samples exceeds the maximum concentration of the standard, the sample diluent can be used to dilute the specimen appropriately and then carry out the experiment. 2. Strictly follow the specified time and temperature for incubation to ensure accurate results. All reagents must be brought to room temperature of 20-25°C before use. keep reagents refrigerated immediately after use. 3. Incorrect plate washing can lead to inaccurate results. Ensure that the wells are as well drained as possible before adding substrate. Do not allow the wells to dry out during incubation. 4. Eliminate liquid residues and fingerprints on the bottom of the plate, otherwise the OD value will be affected. 5. The substrate chromogenic solution should be colorless or very light in color; substrate solution that has turned blue cannot be used. 6. Avoid cross contamination of reagents and specimens to avoid false results. 7. Avoid direct exposure to strong light during storage and incubation. 8. Equilibrate to room temperature before opening the sealed bag to prevent water droplets from condensing on the cold slats. 9. Any reaction reagents should not come into contact with bleaching solvents or strong gases emitted from bleaching solvents. Any bleaching components will destroy the biological activity of the reaction reagents in the kit. 10. Expired products must not be used. 11. If there is a possibility of spreading disease, all samples should be managed and samples and test devices should be handled according to prescribed procedures. |
Coagulation Factor VII (FVII) is a key serine protease in the extrinsic pathway of the blood coagulation cascade, playing a critical role in initiating the coagulation process. It is synthesized in the liver and circulates in the bloodstream as a zymogen, which is activated upon tissue injury to form activated FVII (FVIIa).
Abnormal levels of FVII are closely associated with various pathological conditions, including bleeding disorders, thrombotic diseases, and liver dysfunction. For researchers studying coagulation mechanisms, disease pathogenesis, or drug development targeting the coagulation system, accurate quantification of FVII in biological samples is essential.
ELISA (Enzyme-Linked Immunosorbent Assay) has become the gold standard for quantitative detection of proteins in biological samples due to its high sensitivity, specificity, and reproducibility. The Mouse Coagulation Factor VII (FVII) ELISA Kit is specifically designed to meet the research needs of measuring FVII levels in mouse models, which are widely used in preclinical studies for their genetic similarity to humans and ease of experimental manipulation.
This kit provides a reliable and efficient tool for researchers in fields such as hematology, immunology, pharmacology, and molecular biology, enabling precise analysis of FVII in serum, plasma, tissue homogenates, and other relevant samples to support in-depth research on coagulation-related diseases and therapeutic interventions.
Target-specific design: Specifically recognizes mouse coagulation factor VII (FVII) without cross-reactivity with other soluble structural analogs, ensuring accurate detection results.
Broad sample compatibility: Suitable for in vitro quantitative analysis of FVII in mouse serum, plasma, tissue homogenates, and cell culture supernatants, meeting diverse research sample requirements.
Wide detection range: Covers a concentration range of 0.25 IU/mL to 8 IU/mL, capable of detecting both low and high levels of FVII in samples.
High sensitivity: Has a sensitivity of less than 0.1 IU/mL, enabling the detection of trace amounts of FVII in biological samples.
Excellent reproducibility: Intraplate coefficient of variation (CV) is less than 10% and interplate CV is less than 15%, ensuring consistent and reliable results across experiments.
User-friendly operation: Follows a standard double-antibody one-step sandwich ELISA protocol with clear, step-by-step procedures, suitable for both experienced researchers and laboratory technicians new to ELISA assays.
Stable storage: Shelf life of 6 months when stored at 2-8°C, ensuring product stability and usability within the valid period.
Accurate quantification: Utilizes a highly specific double-antibody sandwich method, minimizing non-specific binding and ensuring precise measurement of FVII concentrations.
Time-efficient workflow: The entire experimental process can be completed within a reasonable timeframe, with incubation steps optimized to balance efficiency and detection performance.
Cost-effective: Provides sufficient reagents for 48 or 96 tests, offering high value for money for laboratories with regular detection needs.
No special equipment required: Only standard laboratory equipment (such as an enzyme labeler, thermostat, and pipettes) is needed, eliminating the need for expensive specialized instruments.
Reliable quality control: Each batch of the kit undergoes strict quality inspection to ensure consistent performance, reducing experimental variability.
Sample-friendly processing: Sample preparation protocols are straightforward, with minimal requirements for sample volume and processing steps, preserving the integrity of FVII in samples.
Research-focused design: Tailored specifically for mouse samples, aligning with the widespread use of mouse models in preclinical coagulation research.
For research use only, not for clinical use.
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