| Field | Specification |
|---|---|
| Target | |
| Alternative names | Coiled-coil domain-containing protein 80; Down-regulated by oncogenes protein 1; Up-regulated in BRS-3 deficient mouse homolog; CCDC80; DRO1; URB; HBE245 |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: NS0; Immunogen sequence: E22-Y950 |
| 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 coiled-coil domain-containing protein 80 (gene CCDC80) in cell culture supernatants, serum and plasma (heparin, EDTA). 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.
CCDC80 (also called URB or DRO1) is a secreted protein that associates with the extracellular matrix. It promotes cell adhesion and matrix assembly, and it has been studied in adipose tissue biology and as a gene whose expression is reduced in some cancers, although its mechanisms are not fully defined.
Kit Components
| Anti-Human CCDC80 Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Human CCDC80 Standard | 2 vials, 10 ng/tube |
| Human CCDC80 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, EDTA) |
| Assay range | 46.9 pg/mL–3,000 pg/mL |
| Sensitivity | <10 pg/mL |
| Standard | Recombinant human CCDC80 (Glu22–Tyr950), expressed in NS0 |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 4.2–6.7% (3 samples, n = 16 each) |
| Inter-assay CV | 5.9–6.8% (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 460 pg/mL.
Example standard curve (OD450, pg/mL): 0 = 0.021; 46.9 = 0.086; 93.8 = 0.145; 187.5 = 0.276; 375 = 0.535; 750 = 1.014; 1500 = 1.701; 3000 = 2.472. 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 | 46.9 | 93.8 | 187.5 | 375 | 750 | 1500 | 3000 |
| O.D. | 0.021 | 0.086 | 0.145 | 0.276 | 0.535 | 1.014 | 1.701 | 2.472 |
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) | 79 | 359 | 1459 | 86 | 438 | 1497 |
| Standard deviation | 3.32 | 24.05 | 80.25 | 5.85 | 25.84 | 91.32 |
| CV (%) | 4.2% | 6.7% | 5.5% | 6.8% | 5.9% | 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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Anwen Yin et al. (2022) Exercise-derived peptide protects against pathological cardiac remodeling. EBioMedicine. 10.1016/j.ebiom.2022.104164
Elena Palà et al. (2022) Proteins and pathways in atrial fibrillation and atrial cardiomyopathy underlying cryptogenic stroke. IJC Heart & Vasculature. 10.1016/j.ijcha.2022.100977
