| Field | Specification |
|---|---|
| Target | |
| Alternative names | C-C motif chemokine 20; Beta-chemokine exodus-1; CC chemokine LARC; CC chemokine ST38; Liver and activation-regulated chemokine; Macrophage inflammatory protein 3 alpha; MIP-3-alpha; Small-inducible cytokine A20; Ccl20; Larc; Scya20; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: E.coli; Immunogen sequence: A28-M97 |
| 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 C-C motif chemokine 20 (gene Ccl20) 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.
CCL20 (also called MIP-3α or LARC) is a C-C chemokine and the only known chemokine ligand of CCR6. It attracts CCR6-expressing immature dendritic cells, memory T cells, Th17 cells and B cells, particularly to epithelial and mucosal surfaces, and it is studied in mucosal immunity, psoriasis and inflammatory bowel disease.
Kit Components
| Rabbit anti-mouse Ccl20 polyclonal antibody (Capture Antibody) | 500 µL, 0.4 mg/mL (recommended dilution 1:100) |
|---|---|
| Biotinylated goat anti-mouse Ccl20 polyclonal antibody (Detection Antibody) | 500 µL, 3.3 µg/mL (recommended dilution 1:100) |
| Avidin-Biotin-Peroxidase Complex (ABC) | 500 µL (recommended dilution 1:100) |
| Lyophilized recombinant mouse Ccl20 standard | 3 × 10 ng/tube |
Kit components are not sold separately.
Performance Data
| Species | Mouse |
|---|---|
| Sample types | Cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA) |
| Assay range | 7.8 pg/mL–500 pg/mL |
| Standard | Recombinant mouse Ccl20 (Ala28–Met97), expressed in E. coli |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 5.5–6.9% (3 samples, n = 16 each) |
| Inter-assay CV | 6.5–8.7% (3 samples, n = 24 each) |
| Format | Reagents for five 96-well plates |
Example standard curve (OD450, pg/mL): 0 = 0.001; 7.8 = 0.09; 15.6 = 0.183; 31.3 = 0.357; 62.5 = 0.644; 125 = 1.136; 250 = 1.78; 500 = 2.179. 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 | 7.8 | 15.6 | 31.3 | 62.5 | 125 | 250 | 500 |
| O.D. | 0.001 | 0.09 | 0.183 | 0.357 | 0.644 | 1.136 | 1.78 | 2.179 |
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) | 9 | 55 | 313 | 9 | 57 | 308 |
| Standard deviation | 0.62 | 3.02 | 21.28 | 0.78 | 3.7 | 21.56 |
| CV (%) | 6.9% | 5.5% | 6.8% | 8.7% | 6.5% | 7% |
Kit components
| Catalog number | Description | Quantity | Storage of opened/reconstituted material |
|---|---|---|---|
| EZ0454-CA | Rabbit anti- mouse Ccl20 polyclonal antibody (Capture Antibody) | 500 µl, 0.4 mg/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. |
| EZ0454-DA | Biotinylated goat anti- mouse Ccl20 polyclonal antibody (Detection Antibody) | 500 µl, 3.3 µg/mL (recommended dilution 1:100) | |
| AR1103 | Avidin-Biotin-Peroxidase Complex (ABC) | 500 µl (recommended dilution 1:100) | |
| EK0454-ST | Lyophilized recombinant mouse Ccl20 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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