| Field | Specification |
|---|---|
| Target | |
| Alternative names | Carboxypeptidase E; CPE; Carboxypeptidase H; CPH; Enkephalin convertase; Prohormone-processing carboxypeptidase |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: NS0; Immunogen sequence: L43-F476 |
| 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 rat carboxypeptidase E (gene Cpe) 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.
Carboxypeptidase E (CPE) is a secretory granule enzyme in neuroendocrine cells that trims C-terminal basic residues from prohormone intermediates to produce active peptides such as insulin and neuropeptides. It also acts as a sorting receptor that directs prohormones into the regulated secretory pathway.
Kit Components
| Anti-Rat Cpe Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Rat Cpe Standard | 2 vials, 10 ng/tube |
| Rat Cpe 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 | Rat |
|---|---|
| Sample types | Cell culture supernatants, serum and plasma (heparin, EDTA) |
| Assay range | 156 pg/mL–10,000 pg/mL |
| Sensitivity | <50 pg/mL |
| Standard | Recombinant rat Cpe (Leu43–Phe476), expressed in NS0 |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 4.6–7.7% (3 samples, n = 16 each) |
| Inter-assay CV | 5.8–6.9% (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 800–1000 pg/mL.
Example standard curve (OD450, pg/mL): 0 = 0.018; 156 = 0.093; 312 = 0.171; 625 = 0.317; 1250 = 0.586; 2500 = 0.945; 5000 = 1.483; 10000 = 1.999. 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. | 0.018 | 0.093 | 0.171 | 0.317 | 0.586 | 0.945 | 1.483 | 1.999 |
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) | 72 | 271 | 858 | 81 | 282 | 926 |
| Standard deviation | 3.96 | 20.87 | 39.47 | 4.70 | 17.48 | 63.89 |
| CV (%) | 5.5% | 7.7% | 4.6% | 5.8% | 6.2% | 6.9% |
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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