- Home
- IVD
- By Technology Types
- By Diseases Types
- By Product Types
- Research
- Resource
- Distributors
- Company
| Product Name | Human Anti-Prothrombinogen IgG Ab (Anti-PT-IgG Ab) ELISA Kit |
| Catalog No. | CFTK-HMM-0072 |
| Test Species | Human |
| Application | This kit is used for the in vitro quantitative analysis of human anti-thrombinogen IgG antibody (Anti-PT-IgG Ab) in serum, plasma, tissue homogenate and related fluid samples. |
| Shelf Life | 6 months |
| Storage | 2-8°C |
| Detection Principle | The kit is an indirect method of enzyme-linked immunosorbent assay (ELISA). To the pre-coated microtiter wells pre-coated with human anti-thrombin IgG antibody (Anti-PT-IgG Ab) capture antigen, add the specimen, standard, and HRP-labeled detection antibody sequentially, and then incubate and wash thoroughly. The color is developed with the substrate TMB, which is converted to blue by catalysis of peroxidase and to final yellow by acid. There is a positive correlation between the shade of color and the human antithrombinogen IgG antibody (Anti-PT-IgG Ab) 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 | 100, 50, 25, 12.5, 6.25, 3.125 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 | 3.125 ng/mL-100 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. |
Prothrombin (coagulation factor II) is a core glycoprotein in the mammalian coagulation cascade, primarily synthesized in the liver and essential for maintaining physiological hemostasis. As an inactive zymogen, it undergoes proteolytic activation by factor Xa (in the presence of cofactors like factor V, calcium ions, and phospholipids) to form thrombin—a key enzyme that catalyzes fibrinogen conversion into insoluble fibrin, the structural backbone of blood clots. Beyond coagulation, prothrombin and its derivatives also participate in immune regulation and inflammatory responses, making them a focal point in interdisciplinary research spanning hematology, immunology, and molecular biology.
The detection of anti-prothrombinogen IgG antibodies (Anti-PT-IgG Ab) has emerged as a critical tool in basic and translational research. These antibodies are known to interact with prothrombin or its cleavage products, potentially disrupting normal coagulation pathways or triggering immune-mediated responses. In research settings, Anti-PT-IgG Ab is widely studied in the context of autoimmune disease models (e.g., antiphospholipid syndrome, systemic lupus erythematosus), thrombotic mechanism investigations, and the development of coagulation-related therapeutics.
Traditional detection methods for Anti-PT-IgG Ab, such as immunoblotting and indirect immunofluorescence, are limited by poor quantitative precision, low throughput, and high variability—hindering large-scale research projects and data reproducibility. The development of ELISA-based detection systems has addressed these challenges, offering a standardized, sensitive, and efficient approach to quantify Anti-PT-IgG Ab in biological samples.
This Human Anti-Prothrombinogen IgG Ab ELISA Kit is specifically engineered for research applications, enabling scientists to accurately measure antibody levels in experimental samples. It supports studies on:
The role of Anti-PT-IgG Ab in coagulation dysfunction and autoimmune pathogenesis.
The correlation between antibody expression and disease progression in animal models.
The evaluation of potential therapeutic agents targeting prothrombin-immune interactions.
High-throughput screening of sample cohorts in epidemiological or preclinical research.
With the growing emphasis on precision medicine and translational research, the demand for reliable, research-grade tools to quantify coagulation-related antibodies continues to rise. This kit is designed to meet the rigorous standards of academic laboratories, biotech companies, and research institutions, providing consistent data to drive scientific discoveries.
Research-Focused Sample Versatility: Supports in vitro quantitative analysis of Anti-PT-IgG Ab in multiple research-grade biological samples, including human serum, plasma (EDTA or heparin-anticoagulated), tissue homogenates, and cell culture supernatants—catering to diverse experimental designs in basic and preclinical research.
Broad Detection Range for Research Flexibility: Offers a detection range of 3.125 ng/mL to 100 ng/mL, covering the expected concentration ranges of Anti-PT-IgG Ab in normal, diseased, or experimentally manipulated research samples.
High Sensitivity for Low-Abundance Detection: Boasts a sensitivity of < 0.1 ng/mL, enabling the detection of low-level Anti-PT-IgG Ab in early-stage experimental models or samples with subtle antibody expression changes—critical for detecting marginal effects in research studies.
Exceptional Specificity for Research Accuracy: Undergoes rigorous validation to ensure no cross-reactivity with other soluble structural analogs (e.g., unrelated coagulation factors, non-specific IgG subtypes, or common serum proteins), guaranteeing that measured signals reflect true Anti-PT-IgG Ab concentrations in research samples.
Reliable Reproducibility for Consistent Data: Maintains intra-plate coefficient of variation < 10% and inter-plate coefficient of variation < 15%, ensuring consistent results across experimental replicates and batches—essential for statistical validity in research studies.
User-Friendly Protocol for Research Efficiency: Features a streamlined workflow with clear, step-by-step instructions, minimizing experimental complexity and reducing hands-on time. The entire assay can be completed within a single working day, supporting high-throughput research projects.
Precise Quantitative Data for Research Validity: Delivers accurate numerical concentrations via a calibrated standard curve (100, 50, 25, 12.5, 6.25, 3.125 ng/mL) and linear regression analysis, providing quantifiable results that can be used for statistical comparisons between experimental groups or longitudinal research tracking.
Cost-Effective Formats for Research Scalability: Available in 48T and 96T formats, allowing laboratories to select the appropriate size based on sample volume—reducing reagent waste for small-scale preliminary studies and lowering per-test costs for large-scale research projects.
Minimal Equipment Dependencies: Only requires a standard enzyme labeler (450 nm wavelength) and basic laboratory consumables (pipettes, centrifuges, 37°C thermostat), which are commonly available in research settings—eliminating the need for expensive specialized equipment and lowering adoption barriers.
Rigorous Quality Control for Research Reliability: Each production batch undergoes comprehensive quality testing, including standard curve linearity, sensitivity, specificity, and reproducibility verification. Only batches meeting strict research-grade standards are released, ensuring consistent performance for critical research experiments.
Research-Centric Technical Support: Accompanied by a detailed datasheet with research-specific guidelines (e.g., tissue homogenization protocols, sample storage recommendations) and troubleshooting tips for common research scenarios. Our technical team is also available to assist with experimental design or result interpretation, supporting successful research outcomes.
Purely Research-Grade Design: Explicitly engineered for research use only, with formulations and validation tailored to the needs of basic science, preclinical, and translational research—ensuring compatibility with research ethics and regulatory requirements for non-clinical applications.
For research use only, not for clinical use.
|
There is no product in your cart. |
Copyright © 2025 Alta DiagenoTech. All rights reserved.
