| Field | Specification |
|---|---|
| Target | |
| Alternative names | Collagen alpha-1 (XVIII) chain;Endostatin;COL18A1; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: E.coli; Immunogen sequence: H1154-S1335 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Serum, Plasma, Milk |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This sandwich ELISA quantifies human collagen alpha-1(XVIII) chain (gene COL18A1) in cell culture supernatants, serum, plasma (heparin, EDTA) and human milk. 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.
Collagen α1(XVIII) chain (COL18A1) is a basement membrane proteoglycan-type collagen important for eye development and neural tube closure. Its C-terminal fragment, endostatin, inhibits endothelial cell migration and angiogenesis, so COL18A1 is studied in vascular biology, retinal development and cancer research.
Kit Components
| Anti-Human COL18A1 Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Human COL18A1 Standard | 2 vials, 10 ng/tube |
| Human COL18A1 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 human milk |
| Assay range | 156 pg/mL–10,000 pg/mL |
| Sensitivity | <10 pg/mL |
| Standard | Recombinant human COL18A1 (His1154–Ser1335), expressed in E. coli |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 4.2–7.2% (3 samples, n = 16 each) |
| Inter-assay CV | 5.7–8.9% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Sample dilution: start at 1:20. In the supplier’s tests, serum and plasma samples measured 30–50 ng/mL.
Example standard curve (OD450, pg/mL): 0 = 0.047; 156 = 0.095; 312 = 0.149; 625 = 0.245; 1250 = 0.432; 2500 = 0.77; 5000 = 1.308; 10000 = 1.98. 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. | 0.047 | 0.095 | 0.149 | 0.245 | 0.432 | 0.77 | 1.308 | 1.98 |
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) | 247 | 1523 | 5144 | 251 | 1462 | 5243 |
| Standard deviation | 14.07 | 109.65 | 216.04 | 14.3 | 130.11 | 319.82 |
| CV (%) | 5.7% | 7.2% | 4.2% | 5.7% | 8.9% | 6.1% |
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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