**Human FVII ELISA Kit – For the Quantitative In Vitro Determination of Human Coagulation Factor VII in Serum, Plasma, Cerebrospinal Fluid, Tissue Homogenate, and Other Body Fluids**
*For Laboratory Research Use Only. Not for Diagnostic or Therapeutic Purposes.*
Before using this product, please read and understand the complete package insert. This ELISA kit is designed for research purposes only and should not be used in clinical diagnostics.
### **INTENDED USE AND TEST PRINCIPLE**
The Human FVII ELISA Kit is intended for the quantitative determination of coagulation factor VII (FVII) in various biological samples such as serum, plasma, cerebrospinal fluid, tissue homogenates, and other body fluids. The test is based on a sandwich ELISA principle, where FVII in the sample binds to specific antibodies coated on the microtiter plate. A horseradish peroxidase (HRP)-conjugated secondary antibody is then added, followed by a chromogenic substrate that produces a color change. The intensity of the color is proportional to the concentration of FVII in the sample. A standard curve is generated using known concentrations of FVII, allowing the quantification of unknown samples.
### **SAMPLE COLLECTION AND STORAGE**
- **Serum:** Collect using a serum separator tube. Allow clotting for 2 hours at room temperature or overnight at 4°C. Centrifuge at 2000×g for 20 minutes. Remove serum and assay immediately or store at -20°C. Avoid repeated freeze-thaw cycles.
- **Plasma:** Use heparin as an anticoagulant. Centrifuge within 30 minutes of collection at 2000×g for 30 minutes at 2–8°C. Store at -20°C.
- **Cell Culture Supernatants, Tissue Homogenates, and Other Biological Fluids:** Centrifuge to remove particulates and assay immediately or store at -20°C. Ensure no hemolysis or granules are present.
### **MATERIALS REQUIRED BUT NOT SUPPLIED**
1. Incubator set at 37°C
2. Microplate reader capable of measuring absorbance at 450 nm
3. Precision pipettes, disposable tips, and absorbent paper
4. Distilled or deionized water
### **REAGENTS PROVIDED**
| Reagent | 96 Determinations | 48 Determinations |
|---------|-------------------|-------------------|
| MicroELISA Strip Plate | 12×8 strips | 12×4 strips |
| Standard (6 vials) | 0.5 ml/vial | 0.5 ml/vial |
| Sample Diluent | 6.0 ml | 3.0 ml |
| HRP-Conjugate Reagent | 10.0 ml | 5.0 ml |
| 20X Wash Solution | 25 ml | 15 ml |
| Chromogen Solution A | 6.0 ml | 3.0 ml |
| Chromogen Solution B | 6.0 ml | 3.0 ml |
| Stop Solution | 6.0 ml | 3.0 ml |
| Closure Plate Membrane | 2 | 2 |
| User Manual | 1 | 1 |
| Sealed Bags | 1 | 1 |
**Note:**
- Standard concentrations: 40, 20, 10, 5, 2.5, 1.25 IU/ml.
- If sample values exceed the highest standard, dilute with Sample Diluent and repeat the assay.
- All reagents must be brought to room temperature before use. Do not thaw samples or reagents in a water bath.
- Do not use expired reagents.
- Use only deionized or distilled water for dilutions.
- Keep microtiter plates in sealed bags until ready for use. Unused strips should be stored at 2–8°C with desiccant.
- Use fresh pipette tips for each transfer to avoid cross-contamination.
### **REAGENT PREPARATION AND STORAGE**
- **Wash Solution (1X):** Dilute 1 volume of 20X Wash Solution with 19 volumes of deionized or distilled water. Store at 2–8°C for up to one month.
### **ASSAY PROCEDURE**
1. Prepare all reagents before starting. Run standards and samples in duplicate.
2. Add 50 µL of standard or sample to appropriate wells. Blank well contains no sample.
3. Add 100 µL of HRP-conjugate reagent to all wells except blank. Cover with adhesive strip and incubate for 60 minutes at 37°C.
4. Wash the plate 4 times.
- **Manual Washing:** Aspirate, fill with 1X Wash Solution, and aspirate again. Repeat four times. Blot dry after final wash.
- **Automated Washing:** Use washer buffer and follow manufacturer instructions.
5. Add 50 µL of Chromogen A and 50 µL of Chromogen B to each well. Incubate for 15 minutes at 37°C, protected from light.
6. Add 50 µL of Stop Solution to each well. Read OD at 450 nm using a microplate reader.
### **DATA ANALYSIS**
- Plot average OD values of standards against their concentrations to generate a standard curve.
- Calculate mean OD for each sample and subtract blank OD.
- Determine FVII concentration by comparing sample OD to the standard curve.
- Intra-assay and inter-assay CV% <15%.
- Assay range: 12.5–40 IU/ml.
- Sensitivity: <1.0 IU/ml.
- No significant cross-reactivity or interference observed.
### **STORAGE AND HANDLING**
- Store at 2–8°C for frequent use; for long-term storage, keep at -20°C.
- Handle all samples and reagents as potentially infectious. Follow proper biosafety protocols.
- Dispose of waste according to local regulations.
- Liquid waste: Add 1.0% sodium hypochlorite and allow 30 minutes for inactivation.
- Substrate solutions may be easily contaminated. Avoid exposure to heat or flame.
This kit is designed for accurate and reliable detection of FVII levels in research settings. Always follow the instructions carefully to ensure consistent and reproducible results.
Lifting Columns
Suzhou Herstar Medical Technology Co., Ltd. , https://www.hosunherstar.com