TWEAK/TNFSF12 Protein, Human/Cynomolgus (HEK293, Fc)

TWEAK Protein refers to the cytokine tumor necrosis factor-like weak inducer of apoptosis, belongs to tumor necrosis factor (TNF) superfamily. TWEAK protein binds to FN14 and TNRFSF12/APO3, is a weak inducer of apoptosis^[1]. TWEAK does have pro-apoptotic activity for tumor cell, mediates NF-kappa-B activation, promotes angiogenesis and the proliferation of endothelial cells (ECs)^[2]. TWEAK also increases IL-6 and IL-8 secretion, and could potentiate the pro-inflammatory activities of TNF and IL-1^[3]. Human TWEAK protein is a type II transmembrane protein (M1-H249) with a transmembrane domain (22-42 a.a.). TWEAK/TNFSF12 Protein, Human/Cynomolgus (HEK293, Fc) is the extracellular part (S97-H249) of TWEAK protein, produced by HEK293 cells with N-terminal mFc-tag.

TWEAK Protein refers to the cytokine tumor necrosis factor-like weak inducer of apoptosis. It is a multifunctional cytokine belonging to tumor necrosis factor (TNF) superfamily, acts function by binding TweakR/Fn14 receptor. TWEAK is a cell surface-associated type II transmembrane protein with 2 types protein chain: the membrane form and the secreted or soluble form. The soluble form derives from the membrane form by proteolytic processing. The protein sequences in human and mouse is very different with similarity of 24.79%^[1].
TWEAK binds to FN14 and possibly also to TNRFSF12/APO3, is a weak inducer of apoptosis in some cell types. TWEAK mediates NF-kappa-B activation, promotes angiogenesis and the proliferation of endothelial cells^[2].
TWEAK has multiple biological activities, many of which are associated with immune system development and function^[1].
TWEAK does have pro-apoptotic activity on a select group of human tumor cell lines and on monocytes, while it promotes cell proliferation in human vascular EC and SMC. Furthermore, FGF-2 co-treatment can potentiate TWEAK-stimulated HUVEC proliferation, an effect that may be due to the ability of FGF-2 to up-regulate TweakR/Fn14 gene expression. At the meanwhile TWEAK-TweakR/Fn14 autocrine signaling promotes human microvascular renal EC (HMREC) migration^[1].
TWEAK also plays key role in inflammatory response. TWEAK, stimulates interleukin (IL)-8 secretion in human tumor cell lines, WI-38 fibroblasts and astrocytes. TWEAK also increases IL-6 secretion and ICAM-1 expression in astrocyte cell. Moreover, TWEAK co-incubation could potentiate the pro-inflammatory activities of TNF and IL-1, and concluded that TWEAK could be involved in the pathogenesis of chronic inflammatory diseases^[3].
Above all, TWEAK involves in stimulation of cell growth and angiogenesis, induction of inflammatory cytokines, and under some experimental conditions, stimulation of apoptosis^[1].

TWEAK (human; 1 ng/mL; changed media 3 times per week for 4 weeks) induces matrix metalloprotease (MMP) production in human chondrocytes^[4].
TWEAK (human; 5 ng/mL; 24 h) and Fn14 interaction induces upregulation of ICAM-1 and E-selectin on HUVEC, also induces chemokine secretion by HUVEC^[5].
TWEAK (human; 1-1 ng/mL; 24-72 h) induces MCP-1 production in HMC in a dose- and time-dependent manner^[6].

Fc-TWEAK (human; 2 μg/mouse; i.p.; twice weekly, dose at , 4, and 7 days) induces inflammatory gene expression and kidney cell proliferation in vivo in C57Bl/6 mice^[6].

1. Measured in a cell proliferation assay using HUVEC human umbilical vein endothelial cells. The ED ₅₀ for this effect is typically 2-8 ng/mL.
2. Immobilized Cynomolgus mFc-TNFSF12 at 10 μg/ml (100 μl/well) can bind human Fc-TNFRSF12A, The EC ₅₀ of human Fc-TNFRSF12A is 0.07-0.15 μg/ml.

• 
  Measured in a cell proliferation assay using HUVEC cells. The ED50 for this effect is 2.745 ng/mL, corresponding to a specific activity is 3.643×10⁵ U/mg.

Cynomolgus

HEK293

N-mFc

F7HGN4 (S94-H249)

407977  [NCBI]

SAPKGRKTRARRAIAAHYEVHPRPGQDGAQAGVDGTVSGWEEARINSSSPLRYNRQIGEFIVTRAGLYYLYCQVHFDEGKAVYLKLDLLVDGVLALRCLEEFSATAASSLGPQLRLCQVSGLLALRPGSSLRIRTLPWAHLKAAPFLTYFGLFQVH

Approximately 34&47 kDa

• 
  Greater than 95% as determined by reducing SDS-PAGE.

Lyophilized powder

Lyophilized from a 0.2 μm filtered solution of 20 mM PB, 150 mM NaCl, pH 7.4.

<1 EU/μg, determined by LAL method.

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH₂O. For long term storage it is recommended to add a carrier protein (0.1% BSA, 5% HSA, 10% FBS or 5% Trehalose).

Stored at -20°C for 2 years. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Room temperature in continental US; may vary elsewhere.

• [1]. Wiley SR, et al. TWEAK, a member of the TNF superfamily, is a multifunctional cytokine that binds the TweakR/Fn14 receptor. Cytokine Growth Factor Rev. 2003 Jun-Aug;14(3-4):241-9.  [Content Brief]

  [2]. Lammens A, et al. Crystal structure of human TWEAK in complex with the Fab fragment of a neutralizing antibody reveals insights into receptor binding. PLoS One. 2013 May 8;8(5):e62697.  [Content Brief]

  [3]. Lynch CN, et al. TWEAK induces angiogenesis and proliferation of endothelial cells. J Biol Chem. 1999 Mar 26;274(13):8455-9.  [Content Brief]

  [4]. Lynch CN, et al. TWEAK induces angiogenesis and proliferation of endothelial cells. J Biol Chem. 1999 Mar 26;274(13):8455-9.  [Content Brief]