| Field | Specification |
|---|---|
| Target | |
| Alternative names | Plasma serine protease inhibitor; Acrosomal serine protease inhibitor; Plasminogen activator inhibitor 3; PAI-3; PAI3; Protein C inhibitor; PCI; Serpin A5; SERPINA5; PCI; PLANH3; PROCI |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: NS0; Immunogen sequence: E26-P406 |
| 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 sandwich ELISA quantifies human plasma serine protease inhibitor (gene SERPINA5) in cell culture supernatants, serum, plasma (heparin, EDTA) and urine. Strips are pre-coated with a capture antibody. After the sample is added, a biotinylated detection antibody binds the captured protein, and an avidin–biotin–peroxidase complex supplies the enzyme. TMB develops color in proportion to the bound target; the reaction is stopped and read at 450 nm, and concentrations are read from the standard curve.
Protein C inhibitor (PCI, SERPINA5) is a heparin-binding serpin found in plasma and many body fluids. It inactivates activated protein C and several other serine proteases, giving it both procoagulant and anticoagulant roles, and it also acts in the male reproductive tract, where it regulates proteases involved in fertilization.
Kit Components
| Anti-Human Serpina5 Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Human Serpina5 Standard | 2 vials, 10 ng/tube |
| Human Serpina5 Biotinylated antibody (100x) | 100 µL |
| Avidin-Biotin-Peroxidase Complex (100x) | 100 µL |
| Sample Diluent | 30 mL |
| Antibody Diluent | 12 mL |
| Avidin-Biotin-Peroxidase Diluent | 12 mL |
| Color Developing Reagent (TMB) | 10 mL |
| Stop Solution | 10 mL |
| Wash Buffer (25x) | 20 mL |
| Adhesive plate sealers | 4 |
Also required (not supplied): microplate reader for 450 nm; incubator; 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) and urine |
| Assay range | 156 pg/mL–10,000 pg/mL |
| Sensitivity | <10 pg/mL |
| Standard | Recombinant human SERPINA5 (Glu26–Pro406), expressed in NS0 |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 7.0–7.9% (3 samples, n = 16 each) |
| Inter-assay CV | 7.6–9.5% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Sample dilution: start at 1:500. In the supplier’s tests, serum and plasma samples measured about 600 ng/mL.
Example standard curve (OD450, pg/mL): 0 = 0.042; 156 = 0.075; 312 = 0.099; 625 = 0.154; 1250 = 0.263; 2500 = 0.522; 5000 = 1.135; 10000 = 2.156. 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 | 156 | 312 | 625 | 1250 | 2500 | 5000 | 10000 |
| O.D. | 0.042 | 0.075 | 0.099 | 0.154 | 0.263 | 0.522 | 1.135 | 2.156 |
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) | 225 | 1716 | 44577 | 220 | 1814 | 45770 |
| Standard deviation | 17.77 | 127.96 | 314.23 | 20.9 | 137.86 | 407.35 |
| CV (%) | 7.9% | 7.3% | 7% | 9.5% | 7.6% | 8.9% |
Kit components
Description|Quantity Pre-coated 96-well strip microplate|1 Standard|2 vials Biotinylated antibody (100x)|100ul Avidin-Biotin-Peroxidase Complex (100x)|100ul Sample Diluent|30ml Antibody Diluent|12ml Avidin-Biotin-Peroxidase Diluent|12ml Color Developing Reagent (TMB)|10ml Stop Solution|10ml Wash Buffer (25x)|20ml Adhesive plate sealers|4Materials required but not provided
- Microplate Reader capable of reading absorbance at 450nm.
- Incubator.
- 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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Song Yunfeng et al. (2022) Therapeutic exosomes loaded with SERPINA5 attenuated endometrial cancer cell migration via the integrin β1/FAK signaling pathway. Cellular Oncology. 10.1007/s13402-022-00687-4
