| Field | Specification |
|---|---|
| Target | |
| Alternative names | Antithrombin-III;ATIII;Serpin C1;SERPINC1;AT3;PRO0309; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: NS0; Immunogen sequence: H33-K464 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Serum, Plasma, Urine |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This one-step sandwich ELISA quantifies human antithrombin-III (gene SERPINC1) in cell culture supernatants, serum, plasma (heparin, EDTA, citrate) and urine. The strips are pre-coated with a capture antibody; sample and an HRP-linked detection antibody are incubated together, so the assay needs fewer wash and incubation steps than a standard sandwich ELISA (the supplier markets the Quick format as a 90-minute assay). TMB develops color in proportion to the bound target, and the plate is read at 450 nm.
Antithrombin-III (antithrombin, SERPINC1) is the main plasma serpin that controls blood coagulation. It inactivates thrombin and factors IXa, Xa and XIa, and its activity is strongly accelerated by heparin, which is the basis of heparin's anticoagulant effect; inherited antithrombin deficiency increases the risk of venous thrombosis.
Kit Components
| 96-well plate precoated with anti-Human SERPINC1 antibody | 1 |
|---|---|
| lyophilized recombinant Human SERPINC1 standard | 20 ng/tube |
| Biotinylated anti-Human SERPINC1 detection antibody (50x) | 100 µL |
| Sample diluent | 30 mL |
| Antibody diluent | 12 mL |
| Wash Buffer (25x) | 20 mL |
| Color Developing Reagent (TMB) | 10 mL |
| Stop Solution | 10 mL |
| Plate Sealers | 4 |
Also required (not supplied): microplate reader for 450 nm; automated plate washer (optional); precision pipettes for 0.5 µL through 1 mL volumes of aqueous solutions; multichannel pipettes (recommended for many samples); deionized or distilled water; 500 mL graduated cylinders; test tubes for dilution.
Kit components are not sold separately.
Performance Data
| Species | Human |
|---|---|
| Sample types | Cell culture supernatants, serum, plasma (heparin, EDTA, citrate) and urine |
| Assay range | 312 pg/mL–20,000 pg/mL |
| Sensitivity | <10 pg/mL |
| Standard | Recombinant human SERPINC1 (His33–Lys464), expressed in NS0 |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 4.2–7.8% (3 samples, n = 16 each) |
| Inter-assay CV | 5.3–8.5% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Example standard curve (OD450, pg/mL): 0 = 0.011; 312 = 0.062; 625 = 0.082; 1,250 = 0.15; 2,500 = 0.295; 5,000 = 0.582; 10,000 = 1.333; 20,000 = 1.974. Run a new standard curve on every plate.
Storage & Handling
Store at 4 °C for up to 6 months or −20 °C for up to 12 months. Avoid repeated freeze–thaw cycles. Ships on gel ice and tolerates up to 3 days at room temperature; freeze on arrival.
Safety
Sample data
| Concentration (pg/ml) | 0 | 312 | 625 | 1,250 | 2,500 | 5,000 | 10,000 | 20,000 |
| O.D. | 0.011 | 0.062 | 0.082 | 0.15 | 0.295 | 0.582 | 1.333 | 1.974 |
Intra/inter assay consistency
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 16 | 16 | 16 | 24 | 24 | 24 |
| Mean (pg/ml) | 647 | 1973 | 8982 | 647 | 2050 | 9365 |
| Standard deviation | 28.46 | 153.89 | 377.24 | 34.29 | 174.25 | 552.53 |
| CV (%) | 4.4% | 7.8% | 4.2% | 5.3% | 8.5% | 5.9% |
Kit components
Description|Quantity 96-well plate|1 lyophilized Standard| Biotinylated detection antibody (50x)|100 ul Avidin-Biotin-Peroxidase Complex (30x)|400 µl Sample diluent|30 ml Antibody diluent|12 ml Avidin-Biotin-Peroxidase Diluent|12 ml Wash Buffer (25x)|20 ml Color Developing Reagent (TMB)|10 ml Stop Solution|10 ml Plate Sealers|4Materials required but not provided
- Microplate Reader capable of reading absorbance at 450nm.
- Automated plate washer (optional).
- Pipettes and pipette tips capable of precisely dispensing 0.5 µl through 1 ml volumes of aqueous solutions.
- Multichannel pipettes are recommended for large amount of samples.
- Deionized or distilled water.
- 500ml graduated cylinders.
- Test tubes for dilution.
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