Rat TNF-alpha ELISA Kit
Cat no : KE20001
Synonyms
Tnf,TNF-alpha,Cachectin,TNF alpha,TNF alpha,TNF-a,
Validation Data Gallery
Product Information
KE20001 is a solid phase sandwich Enzyme Linked-Immuno-Sorbent Assay (Sandwich ELISA). The TNF-α ELISA kit is to be used to detect and quantify protein levels of endogenous TNF-α. The assay recognizes rat TNF-α. An antibody specific for TNF-α has been pre-coated onto the microwells. The TNF-α protein in samples is captured by the coated antibody after incubation. Following extensive washing, another antibody of biotinylated specific for TNF-α is added to detect the captured TNF-α protein. For signal development, Streptavidin-HRP is added, followed by Tetramethyl-benzidine (TMB) reagent. Solution containing sulfuric acid is used to stop color development and the color intensity which is proportional to the quantity of bound protein is measurable at 450 nm with the correction wavelength set at 630 nm.
| Product name | Rat TNF-alpha ELISA Kit |
| Tests | 1 X 96 well plate |
| Sample type | Cell culture supernatants |
| Assay type | Sandwich |
| Sensitivity | 7.7 pg/mL |
| Range | 15.6-1000 pg/mL |
| Reactivity | Rat |
| Tested applications | Sandwich ELISA |
| Gene ID (NCBI) | 24835 |
Recovery
| Sample Type | Average | Range |
|---|---|---|
| Cell culture supernatants | 84% | 71%-94% |
IntraAssay
| Sample | n | mean ( pg/mL) | SD | CV% |
|---|---|---|---|---|
| 1 | 20 | 288.8 | 17.9 | 6.2 |
| 2 | 20 | 598.9 | 40.2 | 6.7 |
| 3 | 20 | 1,088.1 | 79.8 | 7.3 |
InterAssay
| Sample | n | mean ( pg/mL) | SD | CV% |
|---|---|---|---|---|
| 1 | 24 | 238.2 | 16.9 | 7.1 |
| 2 | 24 | 374.0 | 34.2 | 9.2 |
| 3 | 24 | 716.9 | 37.2 | 5.2 |
Background Information
TNF, as also known as TNF-alpha, or cachectin, is a multifunctional proinflammatory cytokine that belongs to the tumor necrosis factor (TNF) superfamily. It is expressed as a 26 kDa membrane bound protein and is then cleaved by TNF-alpha converting enzyme (TACE) to release the soluble 17 kDa monomer, which forms homotrimers in circulation. It is produced chiefly by activated macrophages, although it can be produced by many other cell types such as CD4+ lymphocytes, NK cells, neutrophils, mast cells, eosinophils, and neurons. It can bind to, and thus functions through its receptors TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. This cytokine is involved in the regulation of a wide spectrum of biological processes including cell proliferation, differentiation, apoptosis, lipid metabolism, and coagulation. rat and human TNF-alpha share 79% amino acid sequence identity. Unlike human TNF-alpha, the rat form is glycosylated. In rat deficiency of this gene is associated with defects in response to bacterial infection, with defects in forming organized follicular dendritic cell networks and germinal centers, and with a lack of primary B cell follicles.
Properties
| Storage Instructions | All the reagents are stored at 2-8℃ for 6 months or -20℃ for 12 months. Refer to the protocol for further storage instructions. |
| Synonyms | Tnf,TNF-alpha,Cachectin,TNF alpha,TNF alpha,TNF-a, |
FAQ
1、I have limited sample diluent. How should I handle samples requiring high dilution factors?
Answer: We recommend performing a serial dilution. For example, if human serum requires a 1,000,000-fold dilution, you can use a three-step 100× dilution method as follows:
Step 1: Take 2 uL of human serum and add it to 198 uL of sample diluent, mix thoroughly to obtain a 100-fold diluted sample.
Step 2: Take 2 uL of the 100-fold diluted sample and add it to 198 uL of sample diluent, mix thoroughly to obtain a 10,000-fold diluted sample.
Step 3: Take 2 uL of the 10,000-fold diluted sample and add it to 198 uL of sample diluent, mix thoroughly to obtain a 1,000,000-fold diluted sample.
If further dilution is required, additional serial steps can be used. We do not recommend using any other solutions or water for dilution, except for the sample diluent we suggest. If your sample diluent is still insufficient, please contact our customer service for more sample diluent.
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