Product Pathways - Growth Factors/Cytokines
Mouse Tumor Necrosis Factor-α (mTNF-α) #5178
PhosphoSitePlus® protein, site, and accession data: TNF-a
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Source
Recombinant mouse TNF-α (mTNF-α) Leu80-Leu235 (Accession #NP_038721) was expressed in human 293 cells at Cell Signaling Technology.
Molecular Characterization
Based on amino acid sequencing, greater than 50% of recombinant mTNF-α starts at Leu80 (LRSSS) and has a calculated MW of 17,257. The remainder starts at Ser82 (SSSQN). DTT-reduced and non-reduced protein migrate as 16 kDa polypeptides.
Purity
>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant mTNF-α. All lots are greater than 98% pure.
Bioactivity
The bioactivity of recombinant mTNF-α was determined in a L-929 cell viability assay. The ED50 of each lot is between 2-8 pg/ml.
Coomassie Gel
The purity of recombinant mTNF-α was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant mTNF-α and staining overnight with Coomassie Blue.
Bioactivity
The viability of L-929 cells treated with increasing concentrations of mTNF-α in the presence of 2 ng/ml actinomycin D was assessed. Cells were stained with crystal violet at the end of treatment and the OD595 was determined.
Western Blotting
Western blot analysis of extracts from HeLa cells, untreated or treated with mouse TNF-α for 20 minutes, using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 (upper) and p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 (lower).
Endotoxin
Less than 0.01 ng endotoxin/1 μg mTNF-α.
Formulation
With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 20 μg BSA per 1 μg mTNF-α. Carrier free: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2.
Background
TNF-α, the prototypical member of the TNF protein superfamily, is a homotrimeric type-II membrane protein (1,2). Membrane bound TNF-α is cleaved by the metalloprotease TACE/ADAM17 to generate a soluble homotrimer (2). Both membrane and soluble forms of TNF-α are biologically active. TNF-α is produced by a variety of immune cells including T cells, B cells, NK cells and macrophages (1). Cellular response to TNF-α is mediated through interaction with receptors TNF-R1 and TNF-R2 and results in activation of pathways that favor both cell survival and apoptosis depending on the cell type and biological context. Activation of kinase pathways (including JNK, ERK (p44/42), p38 MAPK and NF-κB) promotes the survival of cells, while TNF-α mediated activation of caspase-8 leads to programmed cell death (1,2). TNF-α plays a key regulatory role in inflammation and host defense against bacterial infection, notably Mycobacterium tuberculosis (3).
- Aggarwal, B.B. (2003) Nat Rev Immunol 3, 745-56.
- Hehlgans, T. and Pfeffer, K. (2005) Immunology 115, 1-20.
- Lin, P.L. et al. (2007) J Investig Dermatol Symp Proc 12, 22-5.
Application References
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Companion Products
- 4370 Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb
- 4695 p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb
- 9872 Phosphate Buffered Saline (PBS-1X) pH7.2 (Sterile)
For Research Use Only. Not For Use In Diagnostic Procedures.
