| Field | Specification |
|---|---|
| Target | |
| Alternative names | Interleukin-27 subunit alpha; IL-27 subunit alpha; IL-27-A; IL27-A; p28; Il27; Il27a |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: NS0; Immunogen sequence: F29-S234 |
| 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 EZ-Set™ kit supplies a matched capture and detection antibody pair and a standard for building your own sandwich ELISA for mouse interleukin-27 subunit alpha (gene Il27) in cell culture supernatants, 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.
IL-27 p28 (IL27, also IL-30) is the subunit that pairs with EBI3 to form the heterodimeric cytokine IL-27. Acting through the WSX-1/gp130 receptor and STAT1/STAT3 signaling, IL-27 has both pro- and anti-inflammatory effects: it favors early Th1 responses, restrains Th2 and Th17 differentiation, and helps limit excessive T-cell activation.
Kit Components
| Mouse anti-mouse Il27 monoclonal antibody (Capture Antibody) | 500 µL, 4 mg/mL (recommended dilution 1:100) |
|---|---|
| Biotinylated goat anti-mouse Il27 polyclonal antibody (Detection Antibody) | 500 µL, 0.15 µg/mL (recommended dilution 1:100) |
| Avidin-Biotin-Peroxidase Complex (ABC) | 500 µL (recommended dilution 1:100) |
| Lyophilized recombinant mouse Il27 standard | 3 × 1 ng/tube |
Kit components are not sold separately.
Performance Data
| Species | Mouse |
|---|---|
| Sample types | Cell culture supernatants, serum and plasma (heparin, EDTA) |
| Assay range | 15.6 pg/mL–1,000 pg/mL |
| Standard | Recombinant mouse Il27 (Phe29–Ser234), expressed in NS0 |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Format | Reagents for five 96-well plates (plates not included) |
Example standard curve (OD450, pg/mL): 0 = 0.008; 15.6 = 0.041; 31.2 = 0.088; 62.5 = 0.164; 125 = 0.298; 250 = 0.579; 500 = 1.037; 1000 = 1.859. 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 | 15.6 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 |
| O.D. | 0.008 | 0.041 | 0.088 | 0.164 | 0.298 | 0.579 | 1.037 | 1.859 |
Kit components
| Catalog number | Description | Quantity | Storage of opened/reconstituted material |
|---|---|---|---|
| EZ0800-CA | Mouse anti- mouse Il27 monoclonal antibody (Capture Antibody) | 500 µl, 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. |
| EZ0800-DA | Biotinylated goat anti- mouse Il27 polyclonal antibody (Detection Antibody) | 500 µl, 0.15 µg/mL (recommended dilution 1:100) | |
| AR1103 | Avidin-Biotin-Peroxidase Complex (ABC) | 500 µl (recommended dilution 1:100) | |
| EK0800-ST | Lyophilized recombinant mouse Il27 standard | 1 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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