Product Pathways - Growth Factors/Cytokines
Human Tumor Necrosis Factor-α (hTNF-α) #8902
Recombinant human TNF-α (hTNF-α) Val77-Leu233 (Accession #HUMTNFAB) was produced in E. coli at Cell Signaling Technology.
Recombinant hTNF-α does not have a Met on the amino terminus and has a calculated MW of 17,352. DTT-reduced and non-reduced protein migrate as 18 kDa polypeptides. The expected amino-terminal VRSSS of recombinant hTNF-α was verified by amino acid sequencing. TNF-α is a non-disulfide-linked homotrimer in solution as determined by chemical cross-linking.
>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hTNF-α. All lots are greater than 98% pure.
The bioactivity of hTNF-α was determined in an L-929 cell viability assay. The ED50 of each lot is between 10-100 pg/ml.
The purity of recombinant hTNF-α was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hTNF-α and staining overnight with Coomassie Blue.
The viability of L-929 cells treated with increasing amounts of hTNF-α in the presence of 2 ng/ml actinomycin D was determined. Cells were stained with crystal violet at the end of treatment and the OD595 was determined.
Western blot analysis of extracts from HeLa cells treated with hTNF-α for 20 minutes, using Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb #3033 (upper) and total NF-κB p65 Antibody #3034 (lower).
Less than 0.01 ng endotoxin/1 μg hTNF-α.
With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 20 μg BSA per 1 μg hTNF-α. Carrier free: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2.
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). The role of TNF-α in autoimmunity is underscored by blocking TNF-α action to treat rheumatoid arthritis and Crohn’s disease (1,2,4).
- 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.
- Brennan, F.M. and McInnes, I.B. (2008) J Clin Invest 118, 3537-45.
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- 4712 TRAF2 Antibody
- 4926 RIP Antibody
- 2682 IKKα Antibody
- 2370 IKKβ (2C8) Rabbit mAb
- 2697 Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb
- 3033 Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb
- 4814 IκBα (L35A5) Mouse mAb (Amino-terminal Antigen)
- 2859 Phospho-IκBα (Ser32) (14D4) Rabbit mAb
- 9243 NF-κB Control Cell Extracts
- 3707 TNF-α Antibody
- 9872 Phosphate Buffered Saline (PBS-1X) pH7.2 (Sterile)
For Research Use Only. Not For Use In Diagnostic Procedures.