| Field | Specification |
|---|---|
| Target | |
| Alternative names | TNF-alpha|APC1 protein; Cachectin; Cachetin; DIF; TNF; TNF, monocyte-derived; TNFA; TNF-A; TNFalpha; TNF-alpha; TNF-alphacachectin; TNFATNF, macrophage-derived; TNFG1F; TNFSF1A; TNFSF2; TNFSF2TNF superfamily, member 2; tumor necrosis factor (TNF superfamily, member 2); tumor necrosis factor alpha; Tumor necrosis factor ligand superfamily member 2; tumor necrosis factor; tumor necrosis factor-alpha |
| 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, Cell Lysate, Serum, Plasma |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This one-step sandwich ELISA quantifies mouse tumor necrosis factor (gene Tnf) in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). The strips are pre-coated with a capture antibody; sample and an HRP-linked detection antibody are incubated together, so the assay needs fewer wash and incubation steps than a standard sandwich ELISA (the supplier markets the Quick format as a 90-minute assay). TMB develops color in proportion to the bound target, and the plate is read at 450 nm.
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
| Anti-Mouse Tnf Pre-coated 96-well Strip Microplate | 12 strips of 8 wells |
|---|---|
| Mouse Tnf Standard | 2 × 10 ng/tube |
| HRP Linked Anti-Mouse Tnf Antibody | 6 mL |
| Sample Diluent | 15 mL |
| TBS-T Wash Buffer (25x) | 12 mL |
| Color Developing Reagent (TMB) | 10 mL |
| Stop Solution | 10 mL |
| Adhesive Plate Sealers | 2 |
Also required (not supplied): microplate reader for 450 nm; automated plate washer (optional); precision pipettes for 0.5 µL through 1 mL volumes of aqueous solutions; multichannel pipettes (recommended for many samples); deionized or distilled water; 500 mL graduated cylinders; test tubes for dilution; horizontal orbital microplate shaker capable of maintaining a speed of 500 rpm, amplitude 3 mm.
Kit components are not sold separately.
Performance Data
| Species | Mouse |
|---|---|
| Sample types | Cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA) |
| Assay range | 15.6 pg/mL–1,000 pg/mL |
| Sensitivity | <1 pg/mL |
| Standard | Recombinant mouse Tnf (Leu80–Leu235), expressed in E. coli |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 5.2–7.5% (3 samples, n = 16 each) |
| Inter-assay CV | 5.9–8.2% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Example standard curve (OD450, pg/mL): 0 = 0.160; 15.6 = 0.232; 31.2 = 0.311; 62.5 = 0.406; 125 = 0.654; 250 = 1.065; 500 = 1.673; 1000 = 2.430. Run a new standard curve on every plate.
Storage & Handling
Store at 4 °C for up to 6 months. Ships on gel ice and tolerates up to 3 days at room temperature; refrigerate on arrival.
Safety
Sample data
| Concentration (pg/ml) | 0 | 15.6 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 |
| O.D. | 0.160 | 0.232 | 0.311 | 0.406 | 0.654 | 1.065 | 1.673 | 2.430 |
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) | 20 | 55 | 314 | 18 | 63 | 350 |
| Standard deviation | 1.04 | 3.74 | 23.55 | 1.06 | 4.59 | 28.7 |
| CV (%) | 5.2% | 6.8% | 7.5% | 5.9% | 7.3% | 8.2% |
Kit components
Description|Quantity 96-well plate|1 lyophilized Standard| Biotinylated detection antibody (50x)|100 ul Avidin-Biotin-Peroxidase Complex (30x)|400 µl Sample diluent|30 ml Antibody diluent|12 ml Avidin-Biotin-Peroxidase Diluent|12 ml Wash Buffer (25x)|20 ml Color Developing Reagent (TMB)|10 ml Stop Solution|10 ml Plate Sealers|4Materials required but not provided
- Microplate Reader capable of reading absorbance at 450nm.
- Automated plate washer (optional).
- Pipettes and pipette tips capable of precisely dispensing 0.5 µl through 1 ml volumes of aqueous solutions.
- Multichannel pipettes are recommended for large amount of samples.
- Deionized or distilled water.
- 500ml graduated cylinders.
- Test tubes for dilution.
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