Cell Signaling Technology

Product Pathways - Growth Factors/Cytokines

Mouse His6 Tumor Necrosis Factor-α (mHis6TNF-α) #4698

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Source

Recombinant mouseHis6 TNF-α (mHis6TNF-α) Leu80-Leu235 (Accession #NP_038721) was expressed in human 293 cells at Cell Signaling Technology.

Molecular Characterization

Recombinant N-terminally His6-tagged mTNF-α has a calculated MW of 20203. DTT-reduced and non-reduced protein migrate as 20 kDa polypeptides.

Purity

>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant mHis6TNF-α. All lots are greater than 98% pure.

Bioactivity

The bioactivity of recombinant mHis6TNF-α was determined in a L-929 cell viability assay. The ED50 of each lot is between 10-40 pg/ml.

Coomassie Gel

Coomassie Gel

The purity of recombinant mHis6TNF-α was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant mHis6TNF-α and staining overnight with Coomassie Blue.

Bioactivity

Bioactivity

The viability of L-929 cells treated with increasing concentrations of mHis6TNF-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 Blotting

Western blot analysis of extracts from HeLa cells, untreated or treated with mHis6TNF-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 mHis6TNF-α.

Formulation

With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 20 μg BSA per 1 μg mHis6TNF-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).

  1. Aggarwal, B.B. (2003) Nat Rev Immunol 3, 745-56.
  2. Hehlgans, T. and Pfeffer, K. (2005) Immunology 115, 1-20.
  3. Lin, P.L. et al. (2007) J Investig Dermatol Symp Proc 12, 22-5.

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For Research Use Only. Not For Use In Diagnostic Procedures.

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