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| Product Name | Human Coagulation Factor Subunit A (FSA) ELISA Kit |
| Catalog No. | CFTK-HMM-0049 |
| Test Species | Human |
| Application | This kit is for the in vitro quantitative analysis of human coagulation factor subunit A (FSA) in serum, plasma, tissue homogenate 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 pre-coated microtiter wells pre-coated with a capture antibody to human coagulation factor subunit A (FSA), the specimen, standard, and HRP-labeled detection antibody are sequentially added, incubated and washed thoroughly. The color is developed with the substrate TMB, which is converted to blue by catalysis of peroxidase and to final yellow by acid. The shade of color is positively correlated with human coagulation factor subunit A (FSA) in the sample. The absorbance (OD value) is measured at 450 nm wavelength using an enzyme meter and the concentration of the sample is calculated. |
| 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 | 16, 8, 4, 2, 1, 0.5 ng/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 | 0.5 ng/mL-16 ng/mL |
| Sensitivity | < 0.1 ng/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 is a complex and precisely regulated biological process that plays a pivotal role in maintaining vascular integrity and preventing excessive bleeding. When blood vessels are damaged, a cascade of coagulation factors is activated sequentially, ultimately leading to the formation of a stable fibrin clot to seal the wound. Among these critical factors, Human Coagulation Factor Subunit A (FSA) stands out as a key component involved in the early stages of the coagulation cascade. Its proper function is essential for initiating the series of reactions that culminate in effective hemostasis.
Dysregulation or abnormalities in FSA expression or activity can have significant clinical and research implications. For instance:
In clinical settings, deficiencies or mutations in FSA may be associated with bleeding disorders, where patients experience prolonged or excessive bleeding even from minor injuries.
Conversely, elevated FSA levels have been linked to an increased risk of thrombotic events, as the overactivation of the coagulation cascade can lead to the formation of unwanted blood clots within blood vessels.
In scientific research, the quantitative detection of FSA is crucial for understanding the underlying mechanisms of coagulation-related diseases, developing targeted therapies, and evaluating the efficacy of potential drug candidates.
The demand for reliable and accurate tools to measure FSA has grown steadily in recent years, driven by advancements in medical research, diagnostic applications, and drug development. ELISA (Enzyme-Linked Immunosorbent Assay) kits have emerged as the gold standard for quantitative protein detection due to their high sensitivity, specificity, and reproducibility.
Exceptional Sensitivity: With a detection limit of less than 0.1 ng/mL, the kit can accurately measure even low concentrations of FSA in samples, ensuring no trace of the target analyte is missed.
Broad Detection Range: Covering a concentration range of 0.5 ng/mL to 16 ng/mL, the kit is versatile enough to handle samples with varying FSA levels, eliminating the need for frequent sample dilution or concentration adjustments.
High Specificity: The kit utilizes highly specific capture and detection antibodies that do not cross-react with other soluble structural analogs, guaranteeing the accuracy and reliability of test results.
Excellent Reproducibility: Boasting an intraplate coefficient of variation (CV) of less than 10% and an interplate CV of less than 15%, the kit delivers consistent results across multiple experiments and different batches, minimizing experimental variability.
User-Friendly Protocol: The one-step sandwich ELISA format simplifies the experimental process, reducing the number of operating steps and saving time. The detailed and clear instructions make it accessible to both experienced researchers and laboratory technicians new to ELISA assays.
Versatile Sample Compatibility: The kit is validated for use with serum, plasma, tissue homogenate, and cell culture supernatant, accommodating a wide range of sample types commonly used in research and clinical studies.
Stable Storage Conditions: With a 6-month shelf life when stored at 2-8°C, the kit offers long-term usability and convenience, allowing laboratories to stock up without concerns about rapid expiration.
Reliable Performance Backed by Rigorous Quality Control: Each batch of the kit undergoes strict quality control testing, including sensitivity, specificity, reproducibility, and stability assessments, ensuring that every unit meets the highest performance standards.
Time and Cost-Efficient: The streamlined experimental procedure reduces the total assay time, enabling researchers to process more samples in less time. Additionally, the kit provides sufficient reagents for 48 or 96 tests, offering cost-effectiveness for both small-scale experiments and large-volume sample analysis.
Accurate and Precise Quantification: The double-antibody one-step sandwich ELISA principle ensures that the signal intensity is directly proportional to the FSA concentration in the sample, enabling precise quantitative results that can be trusted for data analysis and publication.
Minimal Sample Consumption: Only 50 µL of sample is required per test, conserving valuable biological samples that may be difficult or costly to obtain.
No Specialized Equipment Required: The kit is compatible with standard laboratory equipment such as enzyme labelers (450 nm), high-precision pipettes, and 37°C thermostats, eliminating the need for expensive specialized instruments and reducing laboratory setup costs.
Comprehensive Technical Support: Our team of experienced technical experts is available to provide assistance with experimental setup, troubleshooting, and result interpretation, ensuring a smooth and successful assay experience for every customer.
For research use only, not for clinical use.
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