| Field | Specification |
|---|---|
| Target | |
| Alternative names | Interleukin-1 beta; IL-1 beta; Catabolin; IL1B; IL1F2 |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: E.coli; Immunogen sequence: A117-S269 |
| 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 human interleukin-1 beta (gene IL1B) in cell culture supernatants, serum and plasma (heparin, EDTA, citrate). 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-1 beta (IL-1β) is a potent pro-inflammatory cytokine made as an inactive precursor and processed by caspase-1 after inflammasome activation. Acting through IL1R1 and IL1RAP, it induces fever, prostaglandin synthesis, neutrophil recruitment and Th17 differentiation, and it is one of the most studied mediators of inflammation.
Kit Components
| Rabbit anti-human IL1B polyclonal antibody (Capture Antibody) | 500 µL, 0.4 mg/mL (recommended dilution 1:100) |
|---|---|
| Biotinylated goat anti-human IL1B polyclonal antibody (Detection Antibody) | 500 µL, 25 µg/mL (recommended dilution 1:100) |
| Avidin-Biotin-Peroxidase Complex (ABC) | 500 µL (recommended dilution 1:100) |
| Lyophilized recombinant human IL1B standard | 3 × 1 ng/tube |
Kit components are not sold separately.
Performance Data
| Species | Human |
|---|---|
| Sample types | Cell culture supernatants, serum and plasma (heparin, EDTA, citrate) |
| Assay range | 3.9 pg/mL–250 pg/mL |
| Standard | Recombinant human IL1B (Ala117–Ser269), expressed in E. coli |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 4.5–6.1% (3 samples, n = 16 each) |
| Inter-assay CV | 6.3–7.2% (3 samples, n = 24 each) |
| Format | Reagents for five 96-well plates |
Example standard curve (OD450, pg/mL): 0 = 0.022; 3.9 = 0.074; 7.8 = 0.105; 15.6 = 0.249; 31.2 = 0.423; 62.5 = 0.707; 125 = 1.095; 250 = 1.830. 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 | 3.9 | 7.8 | 15.6 | 31.2 | 62.5 | 125 | 250 |
| O.D. | 0.022 | 0.074 | 0.105 | 0.249 | 0.423 | 0.707 | 1.095 | 1.830 |
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) | 23 | 56 | 158 | 26 | 62 | 173 |
| Standard deviation | 1 | 3.4 | 8.5 | 1.6 | 3.9 | 12.4 |
| CV (%) | 4.5% | 6.1% | 5.4% | 6.3% | 6.4% | 7.2% |
Kit components
| Catalog number | Description | Quantity | Storage of opened/reconstituted material |
|---|---|---|---|
| EZ0392-CA | Rabbit anti- human IL1B 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. |
| EZ0392-DA | Biotinylated goat anti- human IL1B polyclonal antibody (Detection Antibody) | 500 µl, 25 µg/mL (recommended dilution 1:100) | |
| AR1103 | Avidin-Biotin-Peroxidase Complex (ABC) | 500 µl (recommended dilution 1:100) | |
| EK0392-ST | Lyophilized recombinant human IL1B 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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