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| Product Name | Human Coagulation Factor XIII A1 (F13A1) ELISA Kit |
| Catalog No. | CFTK-HMM-0038 |
| Test Species | Human |
| Application | This kit is for the in vitro quantitative analysis of Human Coagulation Factor XIII A1 (F13A1) in serum, plasma, tissue homogenates and related fluids. |
| 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 microtiter wells pre-coated with the capture antibody of human coagulation factor XIII A1 (F13A1), 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 XIII A1 (F13A1) in the sample. The absorbance (OD value) is measured at 450 nm wavelength using an enzyme meter and the sample concentration was 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 | 24, 12, 6, 3, 1.5, 0.75 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 | 0.75 IU/mL-24 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 XIII A1 (F13A1), the A chain of Coagulation Factor XIII, is a core component of the human coagulation cascade and fibrinolytic system, with pivotal roles in regulating hemostasis, fibrin clot stabilization, and tissue repair mechanisms. Its unique biological properties and involvement in multiple physiological and pathological pathways have made it a focal point of scientific research, driving the need for reliable tools to quantify F13A1 levels in experimental samples.
Biological Function and Mechanism: F13A1 is predominantly synthesized in megakaryocytes, platelets, and monocytes. Upon activation by thrombin during the coagulation process, it forms a functional heterotetramer with the F13B chain (Factor XIII B chain). This active enzyme acts as a transglutaminase, catalyzing the cross-linking of fibrin monomers through covalent bonds between glutamine and lysine residues. This cross-linking step is critical for enhancing the mechanical stability of fibrin clots, rendering them resistant to fibrinolysis and ensuring effective hemostasis in physiological settings. Beyond coagulation, F13A1 is involved in extracellular matrix remodeling, cell adhesion, and migration—key processes in tissue repair, wound healing, and embryonic development, making it relevant to diverse research fields.
Research Significance Across Disciplines: F13A1 has emerged as a key target in multiple research areas, as its dysregulation is linked to various pathological conditions.
In hematology research, congenital F13A1 deficiency (a rare genetic disorder) is characterized by impaired clot stabilization, leading to recurrent bleeding. Studies on this deficiency rely on accurate F13A1 quantification to explore disease mechanisms, evaluate potential therapeutic interventions, and assess genotype-phenotype correlations.
In cardiovascular research, F13A1 is implicated in thrombus formation and stability. Research has shown that altered F13A1 levels may contribute to hypercoagulable states or increased bleeding risk, making it a valuable marker for studying conditions such as deep vein thrombosis, atherosclerosis, and the effects of anticoagulant drugs in preclinical models.
In oncology and tumor biology, F13A1 is involved in tumor-induced coagulation disorders and the remodeling of the tumor microenvironment. Researchers investigate F13A1 to understand its role in tumor progression, metastasis, and angiogenesis, as well as its potential as a research biomarker for monitoring tumor-related coagulopathy.
In tissue engineering and regenerative medicine, F13A1’s role in wound healing and extracellular matrix formation has sparked interest in its application for developing novel therapeutic strategies for tissue repair and regeneration.
Versatile Sample Compatibility: Validated for quantitative analysis of F13A1 in multiple human research samples, including serum, EDTA/heparin-anticoagulated plasma, tissue homogenates, and cell culture supernatants. This versatility allows researchers to use a single kit for diverse experimental models, eliminating the need for multiple specialized assays.
High Sensitivity for Low-Concentration Samples: With a sensitivity of <0.1 IU/mL, the kit can reliably detect low levels of F13A1—critical for research involving samples from knockout models, F13A1-deficient specimens, or early-stage pathological samples where F13A1 expression is minimal.
Wide Detection Range: Covers a concentration range of 0.75 IU/mL to 24 IU/mL, enabling accurate quantification of both low and high F13A1 levels. This eliminates the need for excessive sample dilution or concentration, simplifying experimental workflows and reducing potential errors.
Exceptional Specificity for Research Accuracy: Utilizes highly specific capture and detection antibodies that do not cross-react with F13B, other coagulation factors (e.g., Factor VIII, Factor IX), or transglutaminase family members. This ensures that measured signals are directly attributed to F13A1, avoiding false results that could compromise research data.
Consistent Reproducibility: Exhibits intra-plate coefficient of variation (CV) <10% and inter-plate CV <15%, ensuring consistent performance across multiple wells and assay runs. Reproducibility is critical for generating reliable, statistically sound data in research studies and facilitating result validation.
User-Friendly Protocol for Research Efficiency: Features a straightforward, step-by-step assay procedure that requires minimal specialized technical expertise. The total assay time is optimized for high throughput, allowing researchers to process multiple samples simultaneously and accelerate experimental timelines.
Stable Storage and Long Shelf Life: Can be stored at 2-8°C for up to 6 months, providing flexibility for research planning and reducing the need for frequent reordering. All components maintain stability under recommended conditions, ensuring consistent performance throughout the kit’s shelf life.
Precise Quantitative Data for Research Insights: Delivers direct, accurate quantification of F13A1 protein concentration, unlike semi-quantitative methods (e.g., Western blotting) or functional assays that measure activity rather than protein levels. This precision enables researchers to compare F13A1 expression across experimental groups, track changes over time, and draw meaningful conclusions about biological processes.
Time and Cost Efficiency for Research Labs: Available in 48T and 96T formats, optimized for high-throughput research. The all-in-one design includes pre-coated microplates and ready-to-use reagents, eliminating the need for researchers to source, validate, or assemble individual components—saving time and reducing experimental costs.
Robust Performance for Challenging Research Samples: Validated to handle common research sample challenges, with clear guidelines for sample processing (e.g., avoiding hemolysis, preventing repeated freeze-thaw cycles). This robustness ensures consistent results even with complex samples such as tissue homogenates or low-quality research specimens.
Research-Grade Quality Compliance: Manufactured to meet strict research standards, with rigorous quality control testing for sensitivity, specificity, and reproducibility. This ensures that the kit consistently delivers reliable data suitable for publication, preclinical studies, and academic research.
Comprehensive Research Support Materials: Includes a detailed datasheet with step-by-step protocols, reagent preparation instructions, troubleshooting tips, and standard curve generation guidance. This supports researchers of all experience levels, reducing the learning curve and minimizing experimental errors.
Focus on Research Flexibility: Compatible with standard laboratory equipment (e.g., 450 nm enzyme labeler, 37°C thermostat) commonly available in research labs, eliminating the need for specialized instrumentation and making the kit accessible to a wide range of research teams.
For research use only, not for clinical use.
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