- Other Names
- Tumor necrosis factor-ž€, Cachectin, Necrosin, Macrophage cytotoxic factor (MCF), Differentiation inducing factor (DIF), TNF superfamily, member 2, (TNFSF2)
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- Product Citations
TNF-α was originally described as an endotoxin-induced, macrophage-derived factor that promotes hemorrhagic necrosis of solid tumors and the cachexia of chronic infections. TNF-α has also been implicated in a range of inflammatory, infectious, and malignant disorders. At the cellular level, TNF-α modulates a broad spectrum of responses including inflammation, immunoregulation, proliferation, apoptosis, and antiviral activity. In bone, the cytokine inhibits extracellular matrix deposition, stimulates matrix metalloprotease synthesis, and enhances production of osteoclastogenic cytokines such as M-CSF and RANKL. Chronic exposure to TNFa in vivo increases osteoclastogenesis through two distinct mechanisms. TNFa first affects osteoclastogenesis at the osteoclast precursor stage in the bone marrow by priming these cells to differentiate into cFms+/CD11b+/RANK+/- osteoclast progenitors via a RANKL/RANK independent mechanism. These osteoclast precursors then enter the blood and peripheral tissues where they differentiate into mature osteoclasts in the presence of RANKL, and this process is accelerated by TNF. The role of TNF at this later stage of osteoclast differentiation is RANKL/RANK dependent. Importantly, TNF-α promotes bone resorption both in vitro and in vivo by enhancing the proliferation and differentiation of osteoclast precursors.Product Details
- Rat TNF-α, amino acids Leu80-Leu235 (Accession # NM_012675) was expressed in E. coli.
- Molecular Mass
- The 157 amino acid N-terminal methionylated recombinant protein has a predicted molecular mass of approximately 17,361 Da. The DTT-reduced protein migrates at approximately 17kDa and the non-reduced protein migrates at approximately 16kDa by SDS-PAGE.
- Purity is >98%, as determined by Coomassie stained SDS-PAGE.
- 0.22µm filtered protein solution is in 10mM NaH2PO4, 150mM NaCl, pH 7.2.
- Endotoxin Level
- Endotoxin level is <0.1 EU/µg (<0.01 ng/µg) protein as determined by the LAL method.
- 10 and 25 µg sizes are bottled at 200 µg/mL. 100 µg size and larger sizes are lot-specific and bottled at the concentration indicated on the vial (please contact technical support for concentration, or use our Lookup tool if you have a lot number.) Please note, new lots of the 100 µg size will be lot-specific and may differ from previous lots that had a fixed concentration.
- Storage & Handling
- Unopened vial can be stored between 2°C and 8°C for one month, at -20°C for six months, or at -70°C for one year. For maximum results, quick spin vial prior to opening. The protein can be aliquoted and stored from -20°C to -70°C. Stock solutions can also be prepared at 50-100 µg/mL in sterile buffer (PBS, HPBS, DPBS, or EBSS) containing carrier protein such as 0.2-1% BSA or HSA and stored in working aliquots at -20°C to -70°C. Avoid repeated freeze/thaw cycles.
- The ED50 is 5- 15 pg/ml, corresponding to a specific activity of 2 - 0.66 x 108units/mg, as determined by a dose dependent cytotoxicity assay using L929 cells treated with actinomycin D.
- Application Notes
BioLegend carrier-free recombinant proteins provided in liquid format are shipped on blue-ice. Our comparison testing data indicates that when handled and stored as recommended, the liquid format has equal or better stability and shelf-life compared to commercially available lyophilized proteins after reconstitution. Our liquid proteins are validated in-house to maintain activity after shipping on blue ice and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at email@example.com.
- TNF superfamily; dimmer/trimmer
- It is produced primarily by macrophages, and it is also be expressed by activated T cells, B cells, NK cells, and neutrophils.
- Type II integral membrane protein processed by TACE for secretion; upregulated by interferons, IL-2, GM-CSF, substance P, bradykinin, PAF, immune complexes, cyclooxygenase; downregulated by IL-6, TGF-β, vitamin D3, prostaglandin E2, PAF antagonists
- Monocytes, neutrophils, macrophages, T cells, fibroblasts, endothelial cells, osteoclasts, adipocytes, astroglia, microglia
- TNFRSF1A (TNF-R1, CD120a, TNFR-p60 Type β, p55); TNFRSF1B (TNF-R2, CD120b, TNFR-p80 Type A, p75)
- Paracrine/endocrine mediator of inflammatory and immune functions; selectively cytotoxic for transformed cells; endothelial cell alterations; chemoattractant
- Biology Area
- Cell Biology, Immunology, Innate Immunity, Neuroinflammation, Neuroscience
- Molecular Family
- Antigen References
1. Boyce BF, et al. J Med 54:127-131 2005.
2. Lam J, et al. J Clin Invest 106:1481-1488 2000.
3. Udagawa N, et al. Arthritis Res4:281-289 2002.
4. Kwon J, et al. Gene 132:227-236 1993.
- Gene ID
- 24835 View all products for this Gene ID
- View information about TNF-alpha on UniProt.org
- Does specific activity of a recombinant protein vary between lots?
Specific activity will vary for each lot and for the type of experiment that is done to validate it, but all passed lots will have activity within the established ED50 range for the product and we guarantee that our products will have lot-to-lot consistency. Please conduct an experiment-specific validation to find the optimal ED50 for your system.
- Have your recombinants been tested for stability?
Our testing shows that the recombinant proteins are able to withstand room temperature for a week without losing activity. In addition the recombinant proteins were also found to withstand four cycles of freeze and thaw without losing activity.
- How do you convert activity as an ED50 in ng/ml to a specific activity in Units/mg?
- Use formula Specific activity (Units/mg) = 10e6/ ED50 (ng/mL)
- How does the activity of your recombinant proteins compare to competitors?
We quality control each and every lot of recombinant protein. Not only do we check its bioactivity, but we also compare it against other commercially available recombinant proteins. We make sure each recombinant protein’s activity is at least as good as or better than the competition’s. In order to provide you with the best possible product, we ensure that our testing process is rigorous and thorough. If you’re curious and eager to make the switch to BioLegend recombinants, contact your sales representative today!
- What is the specific activity or ED50 of my recombinant protein?
The specific activity range of the protein is indicated on the product datasheets. Because the exact activity values on a per unit basis can largely fluctuate depending on a number of factors, including the nature of the assay, cell density, age of cells/passage number, culture media used, and end user technique, the specific activity is best defined as a range and we guarantee the specific activity of all our lots will be within the range indicated on the datasheet. Please note this only applies to recombinants labeled for use in bioassays. ELISA standard recombinant proteins are not recommended for bioassay usage as they are not tested for these applications.