| Field | Specification |
|---|---|
| Target | |
| Alternative names | Interleukin-1 alpha; IL-1 alpha; Il1a |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: E.coli; Immunogen sequence: S115-S270 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Serum |
| 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-1 alpha (gene Il1a) in cell culture supernatants and serum. 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 alpha (IL-1α) is an IL-1 family cytokine stored inside many non-immune cells and released when cells are damaged, acting as an alarmin. It signals through IL1R1 and IL1RAP via MyD88 and IRAK kinases to activate NF-κB and MAPK pathways, and it is studied in sterile inflammation, fever and tissue injury.
Kit Components
| Mouse anti-mouse Il1a monoclonal antibody (Capture Antibody) | 500 µL, 4 mg/mL (recommended dilution 1:100) |
|---|---|
| Biotinylated goat anti-mouse Il1a 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 Il1a standard | 3 × 10 ng/tube |
Kit components are not sold separately.
Performance Data
| Species | Mouse |
|---|---|
| Sample types | Cell culture supernatants and serum |
| Assay range | 4.69 pg/mL–300 pg/mL |
| Standard | Recombinant mouse Il1a (Ser115–Ser270), expressed in E. coli |
| 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 = 0.046; 4.7 = 0.112; 9.4 = 0.142; 18.8 = 0.240; 37.5 = 0.430; 75 = 0.760; 150 = 1.270; 300 = 1.871. 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 | 4.7 | 9.4 | 18.8 | 37.5 | 75 | 150 | 300 |
| O.D. | 0.046 | 0.112 | 0.142 | 0.240 | 0.430 | 0.760 | 1.270 | 1.871 |
Kit components
| Catalog number | Description | Quantity | Storage of opened/reconstituted material |
|---|---|---|---|
| EZ0391-CA | Mouse anti- mouse Il1a 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. |
| EZ0391-DA | Biotinylated goat anti- mouse Il1a 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) | |
| EK0391-ST | Lyophilized recombinant mouse Il1a 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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