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KillerTRAIL™ (soluble) (human), (recombinant)

Enhanced ligand that utilizes the proprietary Killer™ linker peptide that promotes stability.
 
ALX-201-073-C020 20 µg 435.00 USD
 
ALX-201-073-3020 3x20 µg 895.00 USD
Do you need bulk/larger quantities?
 
KillerTRAIL™ (soluble) (human), (recombinant) is an enhanced ligand that utilizes the proprietary Killer™ linker peptide that promotes trimerization to form a more stable oligomer.

It has been well established that signals transmitted via TNF ligands and receptors are critical in mediating inflammation, cancer, and immunity. This has led to drug development aiming to either reduce or enhance receptor response. One limitation in this process is the inherent instability of recombinant ligands and their inability to effectively mimic the natural clustering of TNF receptors on the cell surface required for immune activation. Oligomerization of TNF ligands such as KillerTRAIL™ has been shown to improve stability and significantly enhance immune activation compared to recombinant ligands alone.

Product Specification

Alternative Name:Apo-2L, TNFSF 10, CD253
 
MW:~24kDa (SDS-PAGE)
 
Source:Produced in E. coli. The extracellular domain of human TRAIL (aa 95-281) is fused at the N-terminus to a His-tag and a linker peptide.
 
UniProt ID:P50591
 
Formulation:Liquid. In 20mM HEPES, pH 7.4, containing 150mM sodium chloride, 0.005% Tween 20, 1% sucrose and 1mM DTT.
 
Purity:≥99% (SDS-PAGE, MS analysis)
 
Purity Detail:Affinity purified. Sterile filtered.
 
Endotoxin Content:≤0.01 EU/μg purified protein (LAL test).
 
Species reactivity:Human, Mouse
 
Specificity:Binds to human and mouse TRAIL receptors and osteoprotegerin (OPG).
 
Biological Activity:Induces apoptosis in a concentration range of 10-100ng/ml.
Note: Does not require a cross-linking enhancer for its potent biological activity. For cell lines that require extensive cross-linking of the TRAIL-Rs for killing (e.g. Jurkat) use SuperKillerTRAIL™ (Prod. No. ALX-201-115).
 
Specific Activity:ED50: 20ng/ml (BJAB cells)
 
Reconstitution:For a special storage and dilution buffer see KillerTRAIL™ Storage and Dilution Buffer (Prod. No. ALX-505-005).
 
Shipping:Shipped on Dry Ice
 
Long Term Storage:-80°C
 
Use/Stability:Stable for at least 12 months after receipt when stored at -80° C. 
 
Handling:Once thawed, prepare aliquots and store at -80°C ( please note - to retain full activity the concentration of aliquots should be at least 0.1 mg/mL).  Avoid freeze/thaw cycles. Keep vial closely capped before use, spin the tube at 5000 x g for 30 sec before use.
 
Scientific Background:TNF-related apoptosis-inducing ligand (TRAIL) is a protein ligand that induces the process of cell death called apoptosis.
 
201-073-1
Figure: Schematic structure of human TRAIL (peptide aa 95-281).
201-073-2
Figure: Apoptosis of TRAIL-sensitive cells. Concentration dependence of apoptosis induction in Jurkat and BJAB cells by KillerTRAIL™, Soluble (human) (recombinant) (Prod. No. ALX-201-073) reveals high activity even at concentrations of 10-100ng/ml.Method: 5x104 human Jurkat (JK) or BJAB cells were cultured in 100µl RPMI 1640 medium containing 10% fetal calf serum for 16 hours at 37°C in a 96-well plate in the presence of the indicated concentrations of KillerTRAIL™. Concentrations of KillerTRAIL™ required to induce apoptosis may vary depending on the cell type studied. Cell death was quantified by forward/sideward scatter (FSC/SSC) combined with propidium iodide (PI) staining analysed by flow cytometry.
201-073-3
Figure: Specificity of KillerTRAIL™-induced apoptosis. Concentration dependent blocking of apoptosis induction of BJAB cells using MAb to TRAIL (human) (2E5) (Prod. No. ALX-804-296) reveals specificity of KillerTRAIL™-mediated killing.Method: Human BJAB cells were treated with KillerTRAIL™, Soluble (human) (recombinant) (at 300ng/ml) in the presence or absence of the indicated concentration of the MAb to TRAIL (human) (2E5) or a control IgG1 mouse monoclonal antibody. After 16 hours apoptosis was evaluated by forward/sideward scatter (FSC/SSC) combined with propidium iodide (PI) stain analysed by flow cytometry.
201-073-4
Figure: Apoptosis of TRAIL-sensitive myeloid cell line ML-1.
201-073-5
Figure: Flow cytometry analysis of the TRAIL-sensitive human leukemia cell line HL60 and of TRAIL-resistant K572 cells.
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201-073-1 201-073-2 201-073-3 201-073-4 201-073-5

Product Literature References

Balance between IL-3 and type Iinterferons and their interrelationship with FasL dictates lifespan and effector functions of human basophils: B.R. Hagmann, et al.; Clin. Exp. Allergy (2016), Abstract;
Differences and similarities in TRAIL-and tumor necrosis factor-mediated necroptotic signaling in cancer cells: J. Sosna, et al.; Mol. Cell. Biol. 36, 2626 (2016), Abstract;
The SMAC mimetic BV6 induces cell death and sensitizes different cell lines to TNF-α and TRAIL-induced apoptosis: M. El-Mesery, et al.; Exp. Biol. Med. (2016), Abstract;
Combinatorial treatment of CD95L and gemcitabine in pancreatic Cancer cells induces apoptotic and RIP1-mediated necroptotic cell death network: S. Pietkiewicz, et al.; Exp. Cell Res. 339, 1 (2015), Application(s): Apoptosis induced in pancreatic cancer cells , Abstract;
Sensitization of Melanoma Cells for Death Ligand TRAIL Is Based on Cell Cycle Arrest, ROS Production, and Activation of Proapoptotic Bcl-2 Proteins: S.A. Quast, et al.; J. Invest. Dermatol. 135, 2794 (2015), Abstract;
c-Rel is a critical mediator of NF-κB-dependent TRAIL resistance of pancreatic cancer cells: C. Geismann, et al.; Cell Death Dis. 5, e1455 (2014), Application(s): Apoptosis induced in human pancreatic ductal adenocarcinoma cells (PANC-1 and Patu8998t), Abstract; Full Text
COX-2-independent effects of celecoxib sensitize lymphoma B cells to TRAIL-mediated apoptosis: A.S. Gallouet, et al.; Clin. Cancer Res. 20, 2663 (2014), Abstract;
Intra- and interdimeric caspase-8 self-cleavage controls strength and timing of CD95-induced apoptosis: S.M. Kallenberger, et al.; Sci. Signal. 7, ra23 (2014), Abstract;
Modulation of nuclear factor E2-related factor-2 (Nrf2) activation by the stress response gene immediate early response-3 (IER3) in colonic epithelial cells: a novel mechanism of cellular adaption to inflammatory stress: I. Stachel, et al.; J. Biol. Chem. 289, 1917 (2014), Application(s): Apoptosis induced in human NCM460 colonocytes, Abstract; Full Text
RAF inhibition overcomes resistance to TRAIL-induced apoptosis in melanoma cells: A. Berger, et al.; J. Invest. Dermatol. 134, 430 (2014), Abstract;
TRAF2 inhibits TRAIL- and CD95L-induced apoptosis and necroptosis: I. Karl, et al.; Cell Death Dis. 5, e1444 (2014), Application(s): Cell death induction in primary keratinocytes, HaCaT and HeLa cells, Abstract; Full Text
Aurora and IKK kinases cooperatively interact to protect multiple myeloma cells from Apo2L/TRAIL: L. Mazzera, et al.; Blood 122, 2641 (2013), Application(s): TRAIL signaling analysis in multiple myeloma cells, Abstract;
CD40-directed scFv-TRAIL fusion proteins induce CD40-restricted tumor cell death and activate dendritic cells: M. El-Mesery, et al.; Cell Death Dis. 4, e916 (2013), Application(s): Stimulation of IL-8 production in HeLa and HeLa-CD40 transfected cells, Abstract;
High expression of crystallin αB represents an independent molecular marker for unfavourable ovarian cancer patient outcome and impairs TRAIL- and cisplatin-induced apoptosis in human ovarian cancer cells: J. Volkmann, et al.; Int. J. Cancer 132, 2820 (2013), Application(s): Apoptosis induced in human ovarian cancer cells (OV-MZ-6 and HEY), Abstract; Full Text
ROS-dependent phosphorylation of Bax by wortmannin sensitizes melanoma cells for TRAIL-induced apoptosis: S.A. Quast, et al.; Cell Death Dis. 10, e839 (2013), Abstract; Full Text
General Sensitization of Melanoma Cells for TRAIL-Induced Apoptosis by the Potassium Channel Inhibitor TRAM-34 Depends on Release of SMAC: S.A. Quest, et al.; Plos One 7, e39290 (2012), Abstract; Full Text
Role of Apollon in Human Melanoma Resistance to Antitumor Agents That Activate the Intrinsic or the Extrinsic Apoptosis Pathways: E. Tassi, et al.; Clin. Cancer Res. 18, 3316 (2012), Application(s): Death induction of human melanoma cells, Abstract; Full Text
Efficient melanoma cell killing and reduced melanoma growth in mice by a selective replicating adenovirus armed with tumor necrosis factor-related apoptosis-inducing ligand: L.F. Fecker, et al. ; Hum. Gene Ther. 22, 405 (2011), Abstract;
Inhibition of SREBP1 sensitizes cells to death ligands: Y. Eberhard, et al.; Oncotarget 2, 186 (2011), Abstract; Full Text
Sensitization of melanoma cells for death ligand-induced apoptosis by an indirubin derivative--Enhancement of both extrinsic and intrinsic apoptosis pathways: A. Berger, et al.; Biochem. Pharmacol. 81, 71 (2011), Abstract;
Sensitization of melanoma cells for TRAIL-induced apoptosis by BMS-345541 correlates with altered phosphorylation and activation of Bax: A. Berger, et al.; Biochem. Pharmacol. 81, 71 (2011), Abstract;
Under HEMA conditions, self-replication of human erythroblasts is limited by autophagic death: G. Migliaccio, et al. ; Blood Cells Mol. Dis. 47, 182 (2011), Abstract;
Exploration of the lysis mechanisms of leukaemic blasts by chimaeric T-cells: D. Laurin, et al.; J. Biomed. Biotechnol. 2010, 234540 (2010), Abstract; Full Text
FOXO3A as a key molecule for all-trans retinoic acid-induced granulocytic differentiation and apoptosis in acute promyelocytic leukemia: Y. Sakoe, et al.; Blood 115, 3787 (2010), Application(s): Death induction of NB4 cells, Abstract; Full Text
Human CD34+ cells engineered to express membrane-bound tumor necrosis factor-related apoptosis-inducing ligand target both tumor cells and tumor vasculature: C. Lavazza, et al.; Blood 115, 2231 (2010), Application(s): Multiple myeloma killing studies in mice, Abstract; Full Text
Lipopolysaccharide-induced expression of TRAIL promotes dendritic cell differentiation: Y.S. Cho, et al.; Immunology 130, 504 (2010), Abstract; Full Text
Resistance of cutaneous anaplastic large-cell lymphoma cells to apoptosis by death ligands is enhanced by CD30-mediated overexpression of c-FLIP: F.K. Braun, et al.; J. Invest. Dermatol. 130, 826 (2010), Abstract;
Small molecule inhibition of phosphatidylinositol-3,4,5-triphosphate (PIP3) binding to pleckstrin homology domains: B. Miao, et al.; PNAS 107, 20126 (2010), Application(s): Death induction of sensitized U87MG cells, Abstract; Full Text
HYD1-induced increase in reactive oxygen species leads to autophagy and necrotic cell death in multiple myeloma cells: R.R. Nair, et al.; Mol. Cancer Ther. 8, 2441 (2009), Application(s): Death induction of H929 cells, Abstract; Full Text
IFN-α-Induced Apoptosis in Hepatocellular Carcinoma Involves Promyelocytic Leukemia Protein and TRAIL Independently of p53: K. Herzer, et al.; Cancer Res. 69, 855 (2009), Application(s): Death induction of Hep3B, Huh7, Huh6, HepG2 and Chang cells, Abstract; Full Text
PRMT5, a Novel TRAIL Receptor-Binding Protein, Inhibits TRAIL-Induced Apoptosis via Nuclear Factor-κB Activation: H. Tanaka, et al.; Mol. Cancer Res. 7, 557 (2009), Application(s): Death induction of HeLa, A549, HCT116 and HT1080 cells, Abstract; Full Text
Caspase-8 Cleaves Histone Deacetylase 7 and Abolishes Its Transcription Repressor Function: F.L. Scott, et al.; J. Biol. Chem. 283, 19499 (2008), Application(s): Death induction of HEK293 cells, Abstract; Full Text
Topoisomerase I Requirement for Death Receptor-induced Apoptotic Nuclear Fission: O. Sordet, et al.; J. Biol. Chem. 283, 23200 (2008), Application(s): Death induction of HCT116 and Jurkat cells, Abstract; Full Text
Blockade of death receptor-mediated pathways early in the signaling cascade coincides with distinct apoptosis resistance in cutaneous T-cell lymphoma cells: F.K. Braun, et al.; J. Invest. Dermatol. 127, 2425 (2007), Abstract;
Resistance of melanoma cells to TRAIL does not result from upregulation of antiapoptotic proteins by NF-kappaB but is related to downregulation of initiator caspases and DR4: B.M. Kurbanov, et al.; Oncogene 26, 3364 (2007), Abstract;
Increased death receptor resistance and FLIPshort expression in polycythemia vera erythroid precursor cells: A. Zeuner, et al.; Blood 107, 3495 (2006), Application(s): Death induction of human erythroblasts, Abstract; Full Text
Inorganic selenium sensitizes prostate cancer cells to TRAIL-induced apoptosis through superoxide/p53/Bax-mediated activation of mitochondrial pathway: H. Hu, et al.; Mol. Cancer Ther. 5, 1873 (2006), Application(s): Death induction and sensitation analysis of LNCaP and DU145 cells, Abstract; Full Text
TRAIL promotes metastasis of human pancreatic ductal adenocarcinoma: A. Trauzold, et al.; Oncogene 25, 7434 (2006), Abstract; Full Text
Chronic lymphocytic leukemic cells exhibit apoptotic signaling via TRAIL-R1: M. MacFarlane, et al.; Cell Death Differ. 12, 773 (2005), Abstract;
Telomerase-Dependent Virotherapy Overcomes Resistance of Hepatocellular Carcinomas against Chemotherapy and Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand by Elimination of Mcl-1: T. Wirth, et al.; Cancer Res. 65, 7393 (2005), Application(s): Death induction of sensitized Huh7 and Hep3B cells and Hep3B-derived s.c. tumor xenografts in mice, Abstract; Full Text
Autocrine Secretion of Fas Ligand Shields Tumor Cells from Fas-Mediated Killing by Cytotoxic Lymphocytes: K. Hallermalm, et al.; Cancer Res. 64, 6775 (2004), Application(s): Death induction of Jurkat cells and death induction trials with OCM1 and OCM8 cells, Abstract; Full Text
HPC1/RNASEL Mediates Apoptosis of Prostate Cancer Cells Treated with 2',5'-Oligoadenylates, Topoisomerase I Inhibitors, and Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand: K. Malathi, et al.; Cancer Res. 64, 9144 (2004), Application(s): Death induction of prostate epithelial cells, DU145 and PC3 cells, Abstract; Full Text
The Human Papillomavirus Type 16 E5 Protein Impairs TRAIL- and FasL-Mediated Apoptosis in HaCaT Cells by Different Mechanisms: K. Kabsch & A. Alonso; J. Virol. 76, 12162 (2002), Application(s): Death induction of HaCaT and A31 cells, Abstract; Full Text
TRAIL (Apo2L) suppresses growth of primary human leukemia and myelodysplasia progenitors: M. Plasilova, et al.; Leukemia 16, 67 (2002), Abstract; Full Text
TRAIL and its receptors in the colonic epithelium: a putative role in the defense of viral infections: J. Sträter, et. al.; Gastroenterology 122, 659 (2002), Abstract;
The anti-apoptotic protein BAG-3 is overexpressed in pancreatic cancer and induced by heat stress in pancreatic cancer cell lines: Q. Liao, et al.; FEBS Lett. 503, 151 (2001), Abstract;
The cytokines tumor necrosis factor-α (TNF-α) and TNF-related apoptosis-inducing ligand differentially modulate proliferation and apoptotic pathways in human keratinocytes expressing the human papilloma: J.R. Basile, et al.; J. Biol. Chem. 276, 22522 (2001), Abstract; Full Text

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