| Field | Specification |
|---|---|
| Target | |
| Alternative names | Cadherin-1;ARC-1;Epithelial cadherin;E-cadherin;Uvomorulin;CD324;E-Cad/CTF1;E-Cad/CTF2;E-Cad/CTF3;Cdh1; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: NS0; Immunogen sequence: D157-V709 |
| 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 mouse cadherin-1 (gene Cdh1) 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.
E-cadherin (CDH1) is the calcium-dependent adhesion protein that builds adherens junctions between epithelial cells. Its cytoplasmic tail links to the actin cytoskeleton through catenins, and loss of E-cadherin is a hallmark of epithelial–mesenchymal transition and is well documented in diffuse gastric and lobular breast cancers.
Kit Components
| Anti-Mouse Cdh1 Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Mouse Cdh1 Standard | 2 vials, 10 ng/tube |
| Mouse Cdh1 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 | Mouse |
|---|---|
| Sample types | Cell culture supernatants, serum and plasma (heparin) |
| Assay range | 156 pg/mL–10,000 pg/mL |
| Sensitivity | <15 pg/mL |
| Standard | Recombinant mouse Cdh1 (Asp157–Val709), expressed in NS0 |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 6.0–7.2% (3 samples, n = 16 each) |
| Inter-assay CV | 6.7–8.3% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Sample dilution: start at 1:200. In the supplier’s tests, serum and plasma samples measured 262–339 ng/mL.
Example standard curve (OD450, pg/mL): 0 = 0.044; 156 = 0.072; 312 = 0.104; 625 = 0.162; 1250 = 0.292; 2500 = 0.58; 5000 = 1.126; 10000 = 2.069. 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. | 0.044 | 0.072 | 0.104 | 0.162 | 0.292 | 0.58 | 1.126 | 2.069 |
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) | 242 | 1644 | 5631 | 217 | 1771 | 5410 |
| Standard deviation | 14.52 | 118.36 | 360.38 | 14.53 | 146.99 | 394.93 |
| CV (%) | 6% | 7.2% | 6.4% | 6.7% | 8.3% | 7.3% |
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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Zhiwen Jiang et al. (2017) Preparation and pharmacological evaluation of norcantharidin-conjugated carboxymethyl chitosan in mice bearing hepatocellular carcinoma. Carbohydrate Polymers. 10.1016/j.carbpol.2017.06.072
El-Ashmawy Nahla E. et al. (2014) Biochemical/metabolic changes associated with hepatocellular carcinoma development in mice. Tumor Biology. 10.1007/s13277-014-1714-6
Zhiwen Jiang et al. (2017) Evaluation on biological compatibility of carboxymethyl chitosan as biomaterials for antitumor drug delivery. Journal of Biomaterials Applications. 10.1177/0885328216688337
Gui-Feng Zhuang et al. (2016) Experiment research of cisplatin implants inhibiting transplantation tumor growth and regulating the expression of KLK7 and E-cad of tumor-bearing mice with gastric cancer. Asian Pacific Journal of Tropical Medicine. 10.1016/j.apjtm.2016.04.022
