| Field | Specification |
|---|---|
| Target | |
| Alternative names | Epithelial cell adhesion molecule;Ep-CAM;Adenocarcinoma-associated antigen;Cell surface glycoprotein Trop-1;Epithelial cell surface antigen;Epithelial glycoprotein;EGP;Epithelial glycoprotein 314;EGP314;hEGP314;KS 1/4 antigen;KSA;Major gastrointestinal tumor-associated protein GA733-2;Tumor-associated calcium signal transducer 1;CD326;EPCAM;GA733-2, M1S2, M4S1, MIC18, TACSTD1, TROP1; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: NS0; Immunogen sequence: Q24-K265 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Cell Lysate, Serum, Plasma |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This EZ-Set™ kit supplies a matched capture and detection antibody pair and a standard for building your own sandwich ELISA for human epithelial cell adhesion molecule (gene EPCAM) in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). You coat the plate with the capture antibody, add samples and standards, detect with the biotinylated antibody and an avidin–biotin–peroxidase complex, develop with TMB and read at 450 nm. Plates, diluents, wash buffer, TMB and stop solution are not included.
EpCAM (CD326) is a transmembrane glycoprotein on most epithelial cells that mediates homophilic cell-cell adhesion. It also influences proliferation and stem cell maintenance, and it is highly expressed in many carcinomas. EpCAM is widely used to identify and isolate epithelial and circulating tumor cells, and EPCAM mutations cause congenital tufting enteropathy.
Kit Components
| Mouse anti-human EPCAM/Trop1 monoclonal antibody (Capture Antibody) | 500 µL, 4 µg/mL (recommended dilution 1:100) |
|---|---|
| Biotinylated goat anti-human EPCAM/Trop1 polyclonal antibody (Detection Antibody) | 500 µL (recommended dilution 1:100) |
| Avidin-Biotin-Peroxidase Complex (ABC) | 500 µL (recommended dilution 1:100) |
| Lyophilized recombinant human EPCAM/Trop1 standard | 3 × 10 ng/tube |
Performance Data
| Species | Human |
|---|---|
| Sample types | Cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA) |
| Assay range | 15.6 pg/mL–1,000 pg/mL |
| Standard | Recombinant human EPCAM (Gln24–Lys265), expressed in NS0 |
| Cross-reactivity | None detected against related proteins in supplier testing |
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 | 15.6 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 |
| O.D. | 0.008 | 0.047 | 0.111 | 0.216 | 0.485 | 1.026 | 1.682 | 2.044 |
Kit components
| Catalog number | Description | Quantity | Storage of opened/reconstituted material |
|---|---|---|---|
| EZ0755-CA | Mouse anti-human EPCAM/Trop1 monoclonal antibody (Capture Antibody) | 500 µl, 4 µg/mL (recommended dilution 1:100) | Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. |
| EZ0755-DA | Biotinylated goat anti-human EPCAM/Trop1 polyclonal antibody (Detection Antibody) | 500 µl (recommended dilution 1:100) | |
| AR1103 | Avidin-Biotin-Peroxidase Complex (ABC) | 500 µl (recommended dilution 1:100) | |
| EZ0755-ST | Lyophilized recombinant human EPCAM/Trop1 standard | 10 ng/tube×3 | Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. |
Materials required but not provided
- Microplate reader in standard size.
- Automated plate washer.
- Incubator.
- Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection.
- Clean tubes and Eppendorf tubes.
- 96 well microplate (Cat# AR1100)
- Plate Sealers.
- Capture Antibody Diluent: PBS.
- Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered.
- Substrate Solution: Tetramethylbenzidine (Cat# AR1104)
- Stop Solution: 2 N H2SO4 (Cat# AR1105)
- Wash Buffer (PBS and PBS-T). Recipes:
- PBS: 8g NaCl, 0.2g KCl, 1.15g Na2HPO4, 0.2g KH2PO4, adjust the total volume to 1 L with distilled water, pH 7.2-7.4, 0.2 µm filtered.
- PBS-T: 0.1% Tween® 20 in PBS, pH 7.2-7.4.
- *Item 6 – 12 are included in the EZ Set Accessory Kit (EZA001)
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