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SuperKillerTRAIL™ (soluble) (human), (recombinant)

Enhanced ligand with improved stability providing significantly enhanced immune activation.
 
ALX-201-115-C010 10 µg 435.00 USD
 
ALX-201-115-3010 3x10 µg 922.00 USD
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SuperKillerTRAIL™ (soluble) (human), (recombinant) is an enhanced ligand that utilizes the Killer™ linker peptide mutated to increase disulfide-mediated cross-linking 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 SuperKillerTRAIL™ has been shown to improve stability and significantly enhance immune activation compared to recombinant ligands alone.

Product Specification

Alternative Name:TNFSF 10, Apo-2L, CD253
 
MW:~26kDa
 
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. The active multimeric conformation is stabilized by an inserted mutation allowing an additional CC-bridge.
 
UniProt ID:P50591
 
Concentration:0.5mg/ml
 
Formulation:Liquid. In 20mM HEPES, pH 7.4, containing 150mM sodium chloride, 0.005% Tween 20, 1% sucrose, 1mM DTT.
 
Purity:≥98% (SDS-PAGE). MS analysis, <1% impurity (mainly Hsp70 protein from E. coli)
 
Purity Detail:Affinity purified. Sterile filtered.
 
Endotoxin Content:<0.01EU/µg purified protein (LAL test).
 
Specificity:Binds to human TRAIL receptors.
 
Biological Activity:Induces apoptosis at concentrations of >5ng/ml. Note: Does not require a cross-linking enhancer for its potent biological activity. Shows superior activity on human cell lines that require extensive cross-linking of TRAIL-Rs for killing (e.g. Jurkat).
 
Shipping:Shipped on Dry Ice
 
Long Term Storage:-80°C
 
Use/Stability:Once thawed it is recommended to prepare appropriate aliquots and to store them at -80°C. In order to retain full activity aliquote concentration must be at least 0.1mg/ml; prepare 20µl aliquots and store at -80°C. Working dilution: 2-100ng/ml. Recommended: Prod. No. ALX-505-005, special storage and dilution buffer.
 
Handling:Avoid freeze/thaw cycles.
 
Scientific Background:TNF-related apoptosis-inducing ligand (TRAIL) is a protein ligand that induces the process of cell death called apoptosis.
 
Technical Info/Product Notes:Click here for an example of SuperKillerTRAIL™ used by Eidgenössische Technische Hochschule (ETH) Zürich.
 
201-115n
Figure: Apoptosis of TRAIL-sensitive cells. Apoptosis induction in BJAB cells by SuperKillerTRAIL™, Soluble (human) (rec.) reveals killing activity at concentrations of 10-15ng/ml, on Jurkat cells even as low as 1-10ng/ml. Method: 5x104 human BJAB cells were cultured in 50µl RPMI 1640 medium containing 10% fetal calf serum for 18 hours at 37°C in a 96-well plate in the presence of the indicated concentrations of SuperKillerTRAIL™, Soluble (human) (rec.). Concentrations of SuperKillerTRAIL™, Soluble (human) (rec.) 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 analyzed by flow cytometry.
201-115 3
Figure: Comparison of pro-apoptotic activities of human and mouse SuperKillerTRAIL™, Soluble (human) (rec.) on human tumor cell lines Jurkat and CEM.Method: Cells were treated with mouse and human SuperKillerTRAIL™, Soluble (human) (rec.) (CC-mutant) (50ng/ml) for 14 hours, stained with propidium iodide and analyzed by flow cytometry.
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Product Literature References

Quinacrine mediated sensitization of glioblastoma (GBM) cells to TRAIL through MMP‐sensitive PEG hydrogel carriers: P. Erkoc, et al.; Macromol. Biosci. 17, 1600267 (2017), Abstract;
CRISPR library designer (CLD): software for multispecies design of single guide RNA libraries: F. Heigwer, et al.; Genome Biol. 17, 55 (2016), Abstract; Full Text
Cytoplasmic levels of cFLIP determine a broad susceptibility of breast cancer stem/progenitor-like cells to TRAIL: R. French, et al. ; Mol. Cancer. 14, 209 (2015), Application(s): Death induction of human breast cancer cells, Abstract; Full Text
The cellular apoptosis susceptibility protein (CAS) promotes TRAIL-induced apoptosis and cell proliferation: P. Monian, et al.; J. Biol. Chem. 91, 127 (2015), Application(s): Western blot, Abstract; Full Text
pERK 1/2 inhibit Caspase-8 induced apoptosis in cancer cells by phosphorylating it in a cell cycle specific manner: R. Mandal, et al.; Mol. Oncol. 8, 232 (2014), Application(s): Analysis of human breast and ovarian cancer cells by WB, Abstract;
hnRNP K suppresses apoptosis independent of p53 status by maintaining high levels of endogenous caspase inhibitors: Z. Xiao, et al.; Carcinogenesis 34, 1458 (2013), Application(s): Death induction of hepatocellular carcinoma cells, Abstract;
Survivin Inhibitor YM-155 Sensitizes Tumor Necrosis Factor - Related Apoptosis-Inducing Ligand-Resistant Glioma Cells to Apoptosis through Mcl-1 Downregulation and by Engaging the Mitochondrial Death Pathway: D.R. Premkumar, et al.; J. Pharmacol. Exp. Ther. 346, 201 (2013), Application(s): Death induction of sensitized U373 and LNZ308 glioma cells, Abstract;
TRAIL+ Human Plasmacytoid Dendritic Cells Kill Tumor Cells In Vitro: Mechanisms of Imiquimod- and IFN-α-Mediated Antitumor Reactivity: M.L. Kalb, et al.; J. Immunol. 188, 1583 (2012), Application(s): Death induction of SKMel2, WM793 and Jurkat cells , Abstract; Full Text
Bortezomib Sensitizes Malignant Human Glioma Cells to TRAIL, Mediated by Inhibition of the NF-κB Signaling Pathway: E.P. Jane, et al.; Mol. Cancer Ther. 10, 198 (2011), Application(s): Death induction on a panel of 8 glioma cell lines, Abstract; Full Text
Intrinsic Anticancer Drug Resistance of Malignant Melanoma Cells Is Abrogated by IFN-β and Valproic Acid: W.P. Roos, et al.; Cancer Res. 71, 4150 (2011), Application(s): Death induction of D05, A375, and D14 cells, Abstract; Full Text
Isolation of a TRAIL Antagonist from the Serum of HIV-infected Patients: D.J. Schnepple, et al.; J. Biol. Chem. 286, 35742 (2011), Application(s): Death induction of peripheral blood lymphocytes, Jurkat and HeLa cells, Abstract; Full Text
Sulforaphane targets pancreatic tumor-initiating cells by NF-{kappa}B-induced anti-apoptotic signaling: G. Kallifatidis, et al.; Gut 58, 949 (2009), Abstract;
CD73 Participates in Cellular Multiresistance Program and Protects against TRAIL-Induced Apoptosis: A. Mikhailov, et al.; J. Immunol. 181, 464 (2008), Application(s): Death induction of Jurkat cells, Abstract; Full Text
Hepatitis C Virus Infection Sensitizes Human Hepatocytes to TRAIL-Induced Apoptosis in a Caspase 9-Dependent Manner: L. Lan, et al.; J. Immunol. 181, 4926 (2008), Application(s): Death induction of primary human hepatocytes and Huh7.5 cells, Abstract; Full Text
Resistance to FasL and tumor necrosis factor-related apoptosis-inducing ligand-mediated apoptosis in Sézary syndrome T-cells associated with impaired death receptor and FLICE-inhibitory protein expression: E. Contassot, et al.; Blood 111, 4780 (2008), Application(s): Death induction of CTCL cell lines, SzS and HD cells, Abstract; Full Text
Fas Receptor Clustering and Involvement of the Death Receptor Pathway in Rituximab-Mediated Apoptosis with Concomitant Sensitization of Lymphoma B Cells to Fas-Induced Apoptosis: A.J. Stel, et al.; J. Immunol. 178, 2287 (2007), Application(s): Death induction of Ramos cells , Abstract; Full Text
Human Immunodeficiency Virus Type 1 Protease Cleaves Procaspase 8 In Vivo: Z. Nie, et al.; J. Virol. 81, 6947 (2007), Application(s): Death induction of Jurkat cells, Abstract; Full Text
Coxsackievirus Protein 2BC Blocks Host Cell Apoptosis by Inhibiting Caspase-3: M.A. Salako, et al.; J. Biol. Chem. 281, 16296 (2006), Application(s): Death induction of HeLa cells, Abstract; Full Text
IFNα-stimulated neutrophils and monocytes release a soluble form of TNF-related apoptosis-inducing ligand (TRAIL/Apo-2 ligand) displaying apoptotic activity on leukemic cells: C. Tecchio, et al.; Blood 103, 3837 (2004), Application(s): Death induction of human neutrophils , Abstract; Full Text
Synthetic lethal targeting of MYC by activation of the DR5 death receptor pathway: Y. Wang, et al.; Cancer Cell 5, 501 (2004), Abstract;

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