| Field | Specification |
|---|---|
| Target | |
| Alternative names | CD166 antigen;Activated leukocyte cell adhesion molecule ;BEN ;Protein DM-GRASP ;CD166;Alcam; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: NS0; Immunogen sequence: W28-K527 |
| 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 mouse CD166 antigen (gene Alcam) 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.
ALCAM (CD166) is an immunoglobulin superfamily adhesion molecule that mediates cell-cell contacts both by binding itself and by binding CD6 on T cells. The ALCAM-CD6 interaction supports immunological synapse formation and T cell activation, and ALCAM is also studied in neural development, stem cells and cancer.
Kit Components
| Rat anti-mouse ALCAM/CD166 monoclonal antibody (Capture Antibody) | 500 µL, 4 µg/mL (recommended dilution 1:100) |
|---|---|
| Biotinylated goat anti-mouse ALCAM/CD166 polyclonal antibody (Detection Antibody) | 500 µL (recommended dilution 1:100) |
| Avidin-Biotin-Peroxidase Complex (ABC) | 500 µL (recommended dilution 1:100) |
| Lyophilized recombinant mouse ALCAM/CD166 standard | 3 × 10 ng/tube |
Performance Data
| Species | Mouse |
|---|---|
| Sample types | Cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA) |
| Assay range | 78 pg/mL–5,000 pg/mL |
| Standard | Recombinant mouse Alcam (Trp28–Lys527), 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 | 78 | 156 | 312 | 625 | 1250 | 2500 | 5000 |
| O.D. | 0.066 | 0.098 | 0.164 | 0.294 | 0.544 | 1.007 | 1.821 | 2.388 |
Kit components
| Catalog number | Description | Quantity | Storage of opened/reconstituted material |
|---|---|---|---|
| EZ1184-CA | Rat anti-mouse ALCAM/CD166 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. |
| EZ1184-DA | Biotinylated goat anti-mouse ALCAM/CD166 polyclonal antibody (Detection Antibody) | 500 µl (recommended dilution 1:100) | |
| AR1103 | Avidin-Biotin-Peroxidase Complex (ABC) | 500 µl (recommended dilution 1:100) | |
| EZ1184-ST | Lyophilized recombinant mouse ALCAM/CD166 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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