Speedy™ Mouse TNF-alpha One-Step ELISA Kit
Cat no : SE60082
Synonyms
Tnf, Tnfa, DIF, TNF a, TNF alpha
Validation Data Gallery
Product Information
| Product name | Speedy™ Mouse TNF-alpha One-Step ELISA Kit |
| Tests | 1 X 96 well plate |
| Sample type | Serum, Plasma, Cell culture supernatant |
| Assay type | Sandwich |
| Sensitivity | 0.2 pg/mL |
| Range | 1.95-125 pg/mL |
| Reactivity | Mouse |
| Tested applications | Sandwich ELISA |
| Gene ID (NCBI) | 21926 |
Recovery
| Sample Type | Average | Range |
|---|---|---|
| Human serum | 104% | 90%-116% |
| Cell culture supernatant | 116% | 106%-122% |
IntraAssay
| Sample | n | mean ( pg/mL) | SD | CV% |
|---|---|---|---|---|
| 1 | 8 | 70.6 | 2.1 | 3.0 |
| 2 | 8 | 15.1 | 0.5 | 3.3 |
| 3 | 8 | 7.0 | 0.4 | 5.7 |
InterAssay
| Sample | n | mean ( pg/mL) | SD | CV% |
|---|---|---|---|---|
| 1 | 16 | 68.0 | 3.3 | 4.9 |
| 2 | 16 | 14.8 | 0.6 | 4.1 |
| 3 | 16 | 7.0 | 0.6 | 8.6 |
Background Information
TNF, 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. Mouse and human TNF-alpha share 79% amino acid sequence identity. Unlike human TNF-alpha, the mouse form is glycosylated. In mouse 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, Tnfa, DIF, TNF a, TNF alpha |
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.
2、If thermostatic shaker is not available, will it affect the duration of my experiment and the experimental data?
Answer: If thermostatic shaker is not available, it may take 2.5 hours to complete this experiment. After adding the antibody cocktail, we suggest you incubate the plate at 37℃ for 2 hours. After adding TMB, you only need to incubate the plate at 37℃ for 15 minutes. Through comparative experiments, our R&D personnel have confirmed that you can still obtain valid experimental data, although the signal will be slightly reduced.