| Field | Specification |
|---|---|
| Target | |
| Alternative names | Pancreatic secretory trypsin inhibitor;Serine protease inhibitor Kazal-type 1;Tumor-associated trypsin inhibitor;TATI;SPINK1;PSTI; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: E.coli; Immunogen sequence: D24-C79 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Serum, Plasma |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This sandwich ELISA quantifies human serine protease inhibitor Kazal-type 1 (gene SPINK1) in cell culture supernatants, serum and plasma (heparin). 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.
Serine protease inhibitor Kazal-type 1 (SPINK1, also called PSTI or TATI) is a small secreted trypsin inhibitor. In the pancreas it guards against premature activation of trypsin and other digestive zymogens, and SPINK1 variants are a well-known risk factor for chronic pancreatitis.
Kit Components
| Anti-Human SPINK1 Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Human SPINK1 Standard | 2 vials, 10 ng/tube |
| Human SPINK1 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 and plasma (heparin) |
| Assay range | 156 pg/mL–10,000 pg/mL |
| Sensitivity | <10 pg/mL |
| Standard | Recombinant human SPINK1 (Asp24–Cys79), expressed in E. coli |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 6.9–7.3% (3 samples, n = 16 each) |
| Inter-assay CV | 7.0–8.3% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Sample dilution: start at 1:1. In the supplier’s tests, serum and plasma samples measured about 2 ng/mL.
Example standard curve (OD450, pg/mL): 0 = 0.07; 156 = 0.104; 312 = 0.147; 625 = 0.223; 1250 = 0.394; 2500 = 0.724; 5000 = 1.237; 10000 = 1.9. 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. | 0.07 | 0.104 | 0.147 | 0.223 | 0.394 | 0.724 | 1.237 | 1.9 |
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) | 245 | 1862 | 5817 | 254 | 1784 | 6136 |
| Standard deviation | 16.9 | 137.22 | 418.82 | 17.78 | 148.07 | 468.33 |
| CV (%) | 6.9% | 7.3% | 7.2% | 7% | 8.3% | 7.6% |
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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Mehner Christine et al. (2020) Targeting an autocrine IL-6–SPINK1 signaling axis to suppress metastatic spread in ovarian clear cell carcinoma. Oncogene. 10.1038/s41388-020-01451-4
Christine Mehner et al. (2015) Serine protease inhibitor Kazal type 1 (SPINK1) drives proliferation and anoikis resistance in a subset of ovarian cancers. Oncotarget. 10.18632/oncotarget.5927
Lin Du et al. (2022) Identification of CCL20 and LCN2 as Efficient Serological Tools for Detection of Hepatocellular Carcinoma. Disease Markers. 10.1155/2022/7758735
