| Field | Specification |
|---|---|
| Target | |
| Alternative names | Tumor necrosis factor; Cachectin; TNF-alpha; Tumor necrosis factor ligand superfamily member 2; TNF-a; Tnf; Tnfa; Tnfsf2 |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: E.coli; Immunogen sequence: L80-L235 |
| 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 tumor necrosis factor (gene Tnf) 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.
Tumor necrosis factor (TNF, TNF-α) is a pro-inflammatory cytokine produced mainly by activated macrophages and T cells. Acting through the receptors TNFR1 and TNFR2, it activates NF-κB and MAP kinase pathways and can trigger apoptosis or necroptosis, driving fever and inflammation. TNF is a central target in rheumatoid arthritis and inflammatory bowel disease research and a standard readout in immunology studies.
Kit Components
| Mouse anti-mouse Tnf monoclonal antibody (Capture Antibody) | 500 µL, 4 mg/mL (recommended dilution 1:100) |
|---|---|
| Biotinylated goat anti-mouse Tnf 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 Tnf standard | 3 × 10 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 Tnf (Leu80–Leu235), 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 = 0.051; 15.6 = 0.090; 31.2 = 0.156; 62.5 = 0.281; 125 = 0.443; 250 = 0.723; 500 = 1.284; 1000 = 1.958. 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. | 0.051 | 0.090 | 0.156 | 0.281 | 0.443 | 0.723 | 1.284 | 1.958 |
Kit components
| Catalog number | Description | Quantity | Storage of opened/reconstituted material |
|---|---|---|---|
| EZ0527-CA | Mouse anti- mouse Tnf 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. |
| EZ0527-DA | Biotinylated goat anti- mouse Tnf 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) | |
| EK0527-ST | Lyophilized recombinant mouse Tnf 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)
Can’t Find What You’re Looking For? We can help you source the best match or customize an ELISA solution for your study. Options may include alternative target synonyms, different species reactivity, sample type/matrix compatibility (serum/plasma/lysate/supernatant), assay format (sandwich/competitive), sensitivity/range, detection chemistry (colorimetric/fluorescent/chemiluminescent), plate format (pre-coated/uncoated, strips vs full plate), and bulk or custom packaging. Click Talk to a Scientist to submit a request form, email us at support@biohippo.com, or explore our Research Services for additional support. Our team will be in contact with you shortly.
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