| Field | Specification |
|---|---|
| Target | |
| Alternative names | Tumor necrosis factor;Cachectin;TNF-alpha;Tumor necrosis factor ligand superfamily member 2;TNF-a;Tumor necrosis factor, membrane form;N-terminal fragment;NTF;Intracellular domain 1;ICD1;Intracellular domain 2;ICD2;C-domain 1;C-domain 2;Tumor necrosis factor, soluble form;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, Cell Lysate, Serum, Plasma |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This one-step sandwich ELISA quantifies rat 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-Rat TNF Pre-coated 96-well Strip Microplate | 12 strips of 8 wells |
|---|---|
| Rat TNF Standard | 2 × 10 ng/tube |
| HRP Linked Anti-Rat 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): reader capable of reading absorbance at 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.
Performance Data
| Species | Rat |
|---|---|
| Sample types | Cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA) |
| Assay range | 31.2 pg/mL–2,000 pg/mL |
| Sensitivity | <1 pg/mL |
| Standard | Recombinant rat Tnf (Leu80–Leu235), expressed in E. coli |
| Cross-reactivity | None detected against related proteins in supplier testing |
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 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 | 2000 |
| O.D. | 0.010 | 0.031 | 0.062 | 0.132 | 0.203 | 0.555 | 0.884 | 1.838 |
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) | 54 | 180 | 534 | 36 | 245 | 786 |
| Standard deviation | 3.06 | 11.79 | 33.05 | 2.53 | 10.49 | 52.11 |
| CV (%) | 5.7% | 6.6% | 6.2% | 7.0% | 4.3% | 6.6% |
Kit components
Description|Quantity|Volume Pre-coated 96-well Strip Microplate|1|12 strips of 8 wells Standard||2 HRP-Linked Antibody|1|6ml Sample Diluent|1|15ml TBS-T Wash Buffer (25x)|1|12ml Color Developing Reagent (TMB)|1|10ml Stop Solution|1|10ml Adhesive Plate Sealers|2|PieceMaterials 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.
- Horizontal orbital microplate shaker capable of maintaining a speed of 500 rpm, amplitude 3mm.
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