Product Features
Product Name Human TNF-alpha EasyTest™ ELISA Kit Assay Type Solid Phase Sandwich ELISA Format 96-well strip plate Assay Process One wash step, two incubations Assay Length 1.5 hours Sample Volume Serum , Plasma, 50µL Detection Method Colorimetric Sensitivity <1pg/mL Assay Range 187.5-12000pg/mL Reactivity Human Cross-Reactivity No cross-reactivity observed with available related molecules Gene Names TNF; DIF-alpha; TNFA; TNFSF2; TNLG1F Gene ID 7124 Swissprot ID P01375
Recovery
| Sample Type | Average % Recovery | Range % |
| Serum | 99 | 81-105 |
| Plasma | 98 | 88-110 |
Linearity
| Sample Type | Dilution | % of Expected |
| Serum | 1:2 | 91 |
| 1:4 | 97 | |
| 1:8 | 92 | |
| Plasma | 1:2 | 98 |
| 1:4 | 113 | |
| 1:8 | 105 |
Standard Curve: Human TNF-alpha EasyTest™ ELISA Kit
Kit Components
• Pre-Coated 96-well Strip Microplate • Wash Buffer • Stop Solution • Assay Diluent • Standards • Bioaim human TNF-alpha mix • TMB One-Step Substrate
Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the manual. 2. Add 50 µl of standard or sample to each well. 3. Add 50 µl of Bioaim mix. 4. Incubate 1 h at RT. 5. Wash and add 100 µl of TMB One-Step Substrate Reagent to each well. 6. Incubate 15 min at RT. 7. Add 50 µl of Stop Solution to each well. 8. Read at 450 nm/570 nm.
Background
Tumor necrosis factor alpha (TNF-α), also known as cachectin and TNFSF2, is the prototypic ligand of the TNF superfamily. It is a pleiotropic molecule that plays a central role in inflammation, apoptosis, and immune system development. TNF-α is produced by a wide variety of immune and epithelial cell types. Human TNF-α shares 97% aa sequence identity with rhesus and 71% – 92% with bovine, canine, cotton rat, equine, feline, mouse, porcine, and rat TNF-α. TNF-α trimers bind the ubiquitous TNF RI and the hematopoietic cell-restricted TNF RII, both of which are also expressed as homotrimers. TNF-α regulates lymphoid tissue development through control of apoptosis. It also promotes inflammatory responses by inducing the activation of vascular endothelial cells and macrophages. TNF-α is a key cytokine in the development of several inflammatory disorders. It contributes to the development of type 2 diabetes through its effects on insulin resistance and fatty acid metabolism.
Storage
The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
Regulatory Status
Human TNF-alpha EasyTest™ ELISA Kit is for Research Use Only.
