| Field | Specification |
|---|---|
| Target | |
| Alternative names | C9orf26; DVS27; DVS27 related protein; IL-1F11; IL-33; IL1F11; IL33; IL33_HUMAN; Interleukin-1 family member 11; Interleukin-33; NF-HEV; NFEHEV; Nuclear factor from high endothelial venules; RP11-575C20.2; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: E.coli; Immunogen sequence: S112-T270 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Serum, Plasma, Cell Lysate |
| 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 interleukin-33 (gene IL33) in cell culture supernatants, serum, plasma (heparin, EDTA) and cell lysates. 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.
Interleukin-33 (IL-33) is an IL-1 family cytokine that acts as an “alarmin” released from damaged epithelial and endothelial cells. Binding to its receptor ST2 (IL1RL1) activates NF-κB and MAPK signaling, promoting type 2 immunity through Th2 cells, mast cells, eosinophils and group 2 innate lymphoid cells. Inside the nucleus, full-length IL-33 also associates with chromatin.
Kit Components
| Rabbit anti-human IL33 polyclonal antibody (Capture Antibody) | 500 µL, 0.4 mg/mL (recommended dilution 1:100) |
|---|---|
| Biotinylated goat anti-human IL33 polyclonal antibody (Detection Antibody) | 500 µL, 3.5 µg/mL (recommended dilution 1:100) |
| Avidin-Biotin-Peroxidase Complex (ABC) | 500 µL (recommended dilution 1:100) |
| Lyophilized recombinant human IL33 standard | 3 × 10 ng/tube |
Kit components are not sold separately.
Performance Data
| Species | Human |
|---|---|
| Sample types | Cell culture supernatants, serum, plasma (heparin, EDTA) and cell lysates |
| Assay range | 15.6 pg/mL–1,000 pg/mL |
| Standard | Recombinant human IL33 (Ser112–Thr270), expressed in E. coli |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 5.8–7.8% (3 samples, n = 16 each) |
| Inter-assay CV | 6.3–9.0% (3 samples, n = 24 each) |
| Format | Reagents for five 96-well plates |
Example standard curve (OD450, pg/mL): 0 = 0.029; 15.6 = 0.057; 31.2 = 0.101; 62.5 = 0.189; 125 = 0.335; 250 = 0.632; 500 = 1.152; 1000 = 2.098. 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. | 0.029 | 0.057 | 0.101 | 0.189 | 0.335 | 0.632 | 1.152 | 2.098 |
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) | 30 | 260 | 484 | 28 | 282 | 511 |
| Standard deviation | 1.74 | 16.38 | 37.75 | 1.76 | 22.27 | 45.99 |
| CV (%) | 5.8% | 6.3% | 7.8% | 6.3% | 7.9% | 9% |
Kit components
| Catalog number | Description | Quantity | Storage of opened/reconstituted material |
|---|---|---|---|
| EZ0929-CA | Rabbit anti- human IL33 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. |
| EZ0929-DA | Biotinylated goat anti- human IL33 polyclonal antibody (Detection Antibody) | 500 µl, 3.5 µg/mL (recommended dilution 1:100) | |
| AR1103 | Avidin-Biotin-Peroxidase Complex (ABC) | 500 µl (recommended dilution 1:100) | |
| EK0929-ST | Lyophilized recombinant human IL33 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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Joana Viegas et al. (2024) A Novel Bionebulizer Approach to Study the Effects of Natural Mineral Water on a 3D In Vitro Nasal Model from Allergic Rhinitis Patients. Biomedicines. 10.3390/biomedicines12020408
