Enhanced immune activation compared to other recombinant ligands
Mimics glycosylation of native human FasL
SUPERFASLIGAND® (soluble) (human), (recombinant) features an N-terminus linker shown to improve stability and enhance immune activation compared to recombinant ligands alone.
Produced in HEK 293 cells. The extracellular domain of human FasL (APO-1L; CD95L; CD178) (aa 103-281) is fused at the N-terminus to a linker peptide (26 aa) and a FLAG®-tag. Glycosylation of rhs SUPERFASLIGAND® is similar to natural human FasL.
UniProt ID:
P48023
Concentration:
0.1mg/ml after reconstitution.
Formulation:
Lyophilized. Contains PBS.
Purity:
≥95% (SDS-PAGE)
Endotoxin Content:
<0.1EU/µg purified protein (LAL test; Associates of Cape Cod).
Species reactivity:
Human, Mouse, Rat
Specificity:
Binds to human, mouse and rat Fas (CD95; APO-1).
Biological Activity:
Kills Fas-sensitive cells. Note: Does not require enhancer.
Applications:
ELISA
Application Notes:
ELISA: binds to Fas receptor at 1-100 ng/ml.
Reconstitution:
Reconstitute with 50µl sterile water. Further dilutions should be made with cell culture medium containing 5% fetal calf serum.
Shipping:
Blue Ice
Long Term Storage:
-20°C
Use/Stability:
Stable for at least 6 months after receipt when stored at -20°C.
Handling:
Avoid freeze/thaw cycles. After reconstitution, prepare aliquots and store at -20°C.
Technical Info/Product Notes:
Historical data has shown that SUPERFASLIGAND® kills Fas-sensitive cells at concentrations of >1ng/ml without the use of enhancer. The ED50 has been shown in previous data to be 1ng/ml (A20 cells).
Note: Results using rhsSUPERFASLIGAND® may differ from those obtained with agonistic antibodies!
FLAG is a registered trademark of Sigma-Aldrich Co.
Regulatory Status:
RUO - Research Use Only
SDS-PAGE analysis: Lane 1: MW Marker, Lane 2: 1μg SuperFasL, stained with Imperial stain.
Jurkat Cell Death: 50,000 Jurkat cells per well are incubated with the indicated concentration of SuperFasL for 3 hours. Cell death is determined by Cell-titer AQueous one-solution cell proliferation reagent. Formazan product is allowed to develop for 6 hours before the plate is read at 490nm. % Viability is determined in comparison to control well with no SuperFasL.
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Product Literature References
A non-canonical vitamin K cycle is a potent ferroptosis suppressor: E. Mishima, et al.; Nature 608, 778 (2022), Abstract;
Dusquetide modulates innate immune response through binding to p62: Y. Zhang, et al.; Structure 30, 1055 (2022), Abstract;
Treatment with Mesenchymal Stromal Cells Overexpressing Fas-Ligand Ameliorates Acute Graft-versus-Host Disease in Mice: A.M. Vacaru, et al.; Int. J. Mol. Sci. 23, 534 (2022), Abstract;
Differences in cell-intrinsic inflammatory programs of yolk sac and bone marrow macrophages: S. Elhag, et al.; Cells 10, 3564 (2021), Abstract; Full Text
Primidone blocks RIPK1-driven cell death and inflammation: T. Riebeling, et al.; Cell Death Differ. 28, 1610 (2021), Abstract; Full Text
Regulatory variants in TCF7L2 are associated with thoracic aortic aneurysm: T. Roychowdhury, et al.; Am. J. Hum. Genet. 108, 1578 (2021), Abstract; Full Text
Caspase-1 initiates apoptosis in the absence of gasdermin D: K. Tsuchiya, et al.; Sci. Rep. 10, 2091 (2019), Abstract; Full Text
Circulating Apoptotic Signals During Acute and Chronic Exposure to High Altitude in Kyrgyz Population: D. Kosanovic, et al.; Front. Physiol. 10, 54 (2019), Application(s): Apoptosis stimulation of human PASMCs, Abstract; Full Text
DPS-2: A Novel Dual MEK/ERK and PI3K/AKT Pathway Inhibitor with Powerful Ex Vivo and In Vivo Anticancer Properties: M. Goulielmaki, et al.; Transl. Oncol. 12, 932 (2019), Abstract; Full Text
Fas‐apoptotic inhibitory molecule 2 localizes to the lysosome and facilitates autophagosome‐lysosome fusion through the LC3 interaction region motif–dependent interaction with LC3: C.J. Hong, et al.; FASEB J. 34, 161 (2019), Abstract; Full Text
Systematic genetic mapping of necroptosis identifies SLC39A7 as modulator of death receptor trafficking: A. Fauster, et al.; Cell Death. Differ. 26, 1138 (2019), Abstract;
C-27-carboxylated oleanane triterpenoids up-regulate TRAIL DISC assembly via p38 MAPK and CHOP-mediated DR5 expression in human glioblastoma cells: H.S. Byun, et al.; Biochem. Pharmacol. 158, 243 (2018), Abstract;
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 47, 71 (2017), Abstract;
Doxorubicin induces cardiotoxicity through upregulation of death receptors mediated apoptosis in cardiomyocytes: L. Zhao, et al.; Sci. Rep. 7, 44735 (2017), Abstract; Full Text
Pevonedistat, a Nedd8-activating enzyme inhibitor, sensitizes neoplastic B-cells to death receptor-mediated apoptosis: C. Paiva, et al.; Oncotarget 8, 21128 (2017), Abstract; Full Text
Regulation of human glioblastoma cell death by combined treatment of cannabidiol, γ-radiation and small molecule inhibitors of cell signaling pathways: V.N. Ivanov, et al.; Oncotarget 8, 74068 (2017), Abstract; Full Text
Antiapoptotic Effect of Acetylcholine in Fas-Induced Apoptosis in Human Keratocytes: M. Sioniecka, et al.; Invest. Ophthalmol. Vis. Sci. 57, 5892 (2016), Abstract;
Desensitization of idiopathic pulmonary fibrosis fibroblasts to Alternaria alternata extract-mediated necrotic cell death: J. Im, et al.; Physiol. Rep. 4, e13020 (2016), Abstract; Full Text
IAP antagonists Birinapant and AT-406 efficiently synergise with either TRAIL, BRAF, or BCL-2 inhibitors to sensitise BRAFV600E colorectal tumour cells to apoptosis: P. Perimenis, et al.; BMC Cancer 16, 624 (2016), Application(s): Apoptosis assays, Abstract; Full Text
Idiopathic pulmonary fibrosis fibroblasts become resistant to Fas ligand-dependent apoptosis via the alteration of decoy receptor 3: J. Im, et al.; J. Pathol. 240, 25 (2016), Application(s): Apoptosis assays on collagen, Abstract;
Microfluidic single-cell transcriptional analysis rationally identifies novel surface marker profiles to enhance cell-based therapies: R. Rennert, et al.; Nat. Commun. 7, 11945 (2016), Application(s): Apoptosis in in-vitro survival assays, Abstract; Full Text
Molecular architecture of the DED chains at the DISC: regulation of procaspase-8 activation by short DED proteins c-FLIP and procaspase-8 prodomain: K. Schleich, et al.; Cell Death Differ. 23, 681 (2016), Application(s): Flow cytometry and cell viability, Abstract;
A cellular screen identifies ponatinib and pazopanib as inhibitors of necroptosis: A. Fauster, et al.; Cell Death Dis. 6, e1767 (2015), Application(s): Cell viability assays, Abstract; Full Text
Brazilin Limits Inflammatory Responses through Induction of Prosurvival Autophagy in Rheumatoid Fibroblast-Like Synoviocytes: H. Lee, et al.; PLoS One 10, e0136122 (2015), Application(s): Apoptosis assays, Abstract; Full Text
IL22/IL-22R Pathway Induces Cell Survival in Human Glioblastoma Cells: H. Akil, et al.; PLoS One 10, e0119872 (2015), Application(s): Cell Culture , Abstract; Full Text
LFA-1-targeting Leukotoxin (LtxA; Leukothera®) causes lymphoma tumor regression in a humanized mouse model and requires caspase-8 and Fas to kill malignant lymphocytes: K. M. DiFranco, et al.; Leuk. Res. 39, 649 (2015), Abstract;
Mitochondrial division inhibitor 1 (mdivi-1) enhances death receptor-mediated apoptosis in human ovarian cancer cells: J. Wang, et al.; Biochem. Biophys. Res. Commun. 456, 7 (2015), Application(s): Death induction in A2780 ovarian cancer cells, Abstract; Full Text
Quantification of apoptosis and necroptosis at the single cell level by a combination of Imaging Flow Cytometry with classical Annexin V/propidium iodide staining: S. Pietkiewicz, et al.; J. Immunol. Methods 423, 99 (2015), Application(s): Death induction in Jurkat cells, Abstract;
The caspase inhibitor zVAD increases lung inflammation in pneumovirus infection in mice: E. van den Berg, et al.; Physiol. Rep. 3, e12332 (2015), Application(s): In vivo injection in mouse, Abstract; Full Text
A novel homozygous Fas ligand mutation leads to early protein truncation, abrogation of death receptor and reverse signaling and a severe form of the autoimmune lymphoproliferative syndrome: S. Nabhani, et al.; Clin. Immunol. 155, 231 (2014), Abstract;
Bacteria induce prolonged PMN survival via a phosphatidylcholine-specific phospholipase C-and protein kinase C-dependent mechanism: S.F. Erttmann, et al.; PLoS One. 9, e87859 (2014), Abstract; Full Text
Lyapunov exponents and phase diagrams reveal multi-factorial control over TRAIL-induced apoptosis: B.B. Aldridge, et al.; Mol. Syst. Biol. 7, 553 (2014), Application(s): Death induction of HCT116, T47D and SKW6.4 cells, Abstract; Full Text
FAIM-L Is an IAP-Binding Protein That Inhibits XIAP Ubiquitinylation and Protects from Fas-Induced Apoptosis: R.S. Moubarak, et al.; J. Neurosci. 33, 19262 (2013), Application(s): Death induction of rat neuronal Type II cells and murine cortical neurons, Abstract;
Low FasL levels promote proliferation of human bone marrow-derived mesenchymal stem cells, higher levels inhibit their differentiation into adipocytes: M.R. Rippo, et al.; Cell Death Dis. 4, e594 (2013), Application(s): Induction of apoptosis or proliferation in human bone marrow mesenchymal stem cell (BM–MSC) , Abstract;
Nucleolin inhibits Fas ligand binding and suppresses Fas-mediated apoptosis in vivo via a surface nucleolin-Fas complex: J.F. Wise, et al.; Blood 121, 4729 (2013), Application(s): Apoptosis induction in Raji, Jurkat, and BC-3 cell lines, Abstract; Full Text
Resistin-like molecule α stimulates proliferation of mesenchymal stem cells while maintaining their multipotency: I.A. Kolosova,et al.; Stem Cells Dev. 22, 239 (2013), Application(s): Apoptosis induction in mesenchymal stem cells, Abstract; Full Text
Roscovitine sensitizes leukemia and lymphoma cells to tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis: J. Molinsky, et al.; Leuk. Lymphoma. 54, 372 (2013), Abstract;
Surface tethered epidermal growth factor protects proliferating and differentiating multipotential stromal cells from FasL-induced apoptosis: M. Rodrigues, et al.; Stem Cells 31, 104 (2013), Application(s): Apoptosis assays, Abstract; Full Text
Fas Ligand-Fas Signaling Participates in Light-Induced Apoptotic Death in Photoreceptor Cells: Q. Chang, et al.; Invest. Ophthalmol. Vis. Sci. 53, 3703 (2012), Application(s): Death induction of 661W cells, Abstract; Full Text
Is Toso an antiapoptotic protein or an Fc receptor for IgM?: K. Honjo, et al.; Blood 119, 1789 (2012), Application(s): Death induction of Jurkat cells, Abstract; Full Text
MS-275 sensitizes osteosarcoma cells to Fas ligand-induced cell death by increasing the localization of Fas in membrane lipid rafts: K. Rao-Bindal, et al.; Cell Death Dis. 3, e369 (2012), Application(s): Apoptosis induction in MS-275 sensitized osteosarcoma cells, Abstract; Full Text
Production of Reactive Oxygen Species by Multipotent Stromal Cells/Mesenchymal Stem Cells Upon Exposure to Fas Ligand: M. Rodrigues, et al; Cell Transplant. 21, 2171 (2012), Application(s): Reactive oxygen species assay with Multipotent stromal cells, Abstract; Full Text
Antioxidant c-FLIP Inhibits Fas Ligand-Induced NF-κB Activation in a Phosphatidylinositol 3-Kinase/Akt-Dependent Manner: A.K. Iyer, et al.; J. Immunol. 187, 3256 (2011), Application(s): Death induction and NF-κB reporter gene assays with 293T and Jurkat cells, Abstract; Full Text
Distinct TRAIL resistance mechanisms can be overcome by proteasome inhibition but not generally by synergizing agents: C. Menke, et al.; Cancer Res. 71, 1883 (2011), Application(s): BJAB cells used in cell death assays with SuperFas Ligand , Abstract; Full Text
TRAIL Receptor Signaling Regulation of Chemosensitivity In Vivo but Not In Vitro: C. Menke, et al.; PLoS One 6, e14527 (2011), Application(s): BJAB cells used in cell death assays with SuperFas Ligand , Abstract; Full Text
A new function of the Fas-FasL pathway in macrophage activation: R. Chakour, et al.; J. Leukoc. Biol. 86, 81 (2009), Application(s): Death induction of bone marrow-derived macrophages and A20B lymphoma cells, Abstract; Full Text
Fas Death Receptor Enhances Endocytic Membrane Traffic Converging into the Golgi Region: M. Degli Esposti, et al.; Mol. Biol. Cell 20, 600 (2009), Application(s): Studies on endocytosis and intracellular trafficking with Jurkat cells, Abstract; Full Text
Regulation of Fas-mediated immune homeostasis by an activation-induced protein, Cyclon: S. Saint Fleur, et al.; Blood 114, 1355 (2009), Application(s): Death induction of CD4+ and CD8+ T cells, Abstract; Full Text
Fas-mediated killing of primary prostate cancer cells is increased by mitoxantrone and docetaxel: J.C. Symes, et al.; Mol. Cancer Ther. 7, 3018 (2008), Application(s): Death induction of primary prostate tumor cultures, 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
The Fas Death Signaling Pathway Connecting Reactive Oxygen Species Generation and FLICE Inhibitory Protein Down-Regulation: L. Wang, et al.; J. Immunol. 180, 3072 (2008), Application(s): Death induction of AE2 and A549 cells, Abstract; Full Text
A role of TRAIL in killing osteoblasts by myeloma cells: I. Tinhofer, et al.; FASEB J. 20, 759 (2006), Application(s): Death induction of primary osteoblasts, Abstract; Full Text
Human Tumor-Released Microvesicles Promote the Differentiation of Myeloid Cells with Transforming Growth Factor-β-Mediated Suppressive Activity on T Lymphocytes: R. Valenti R., et al.; Cancer Res. 66, 9290 (2006), Application(s): Death induction of CD14+ monocytes and Jurkat cells, Abstract; Full Text
Integrated mechanistic and data-driven modelling for multivariate analysis of signalling pathways: F. Hua, et al.; J. R. Soc. Interface 3, 515 (2006), Application(s): Caspase-8 and -3 cleavage kinetics in Jurkat cells, Abstract; Full Text
Phosphatidylinositol 3-Kinase/Akt Positively Regulates Fas (CD95)-Mediated Apoptosis in Epidermal Cl41 Cells: B. Lu, et al.; J. Immunol. 176, 6785 (2006), Application(s): Death induction, PI3K/Akt phosphorylation assays, ELISA-based DNA fragmentation, ROS production and Fas promoter activity assays in Cl41 cells, Abstract; Full Text
Effects of Bcl-2 Levels on Fas Signaling-Induced Caspase-3 Activation: Molecular Genetic Tests of Computational Model Predictions: F. Hua, et al.; J. Immunol. 175, 985 (2005), Application(s): Death induction of Jurkat cells, Abstract; Full Text
Haploinsufficiency, rather than the effect of an excessive production of soluble CD95 (CD95ΔTM), is the basis for ALPS Ia in a family with duplicated 3' splice site AG in CD95 intron 5 on one allele: J. Roesler, et al.; Blood 106, 1652 (2005), Application(s): Death induction of Jurkat cells and normal activated peripheral T cells, Abstract; Full Text
In vitro engagement of CD3 and CD28 corrects T cell defects in chronic lymphocytic leukemia: M. Bonyhadi, et al.; J. Immunol. 174, 2366 (2005), Abstract; Full Text
Nitric Oxide Negatively Regulates Fas CD95-induced Apoptosis through Inhibition of Ubiquitin-Proteasome-mediated Degradation of FLICE Inhibitory Protein: P. Chanvorachote, et al.; J. Biol. Chem. 280, 42044 (2005), Application(s): Death induction and analysis of regulation of Fas-induced apoptosis in BEAS-2B cells, Abstract; Full Text
Fas Ligand Down-Regulates Cytokine-Induced Fas Receptor Expression on Insulinoma (NIT-1), But Not Islet Cells, from Autoimmune Nonobese Diabetic Mice: P. Augstein, et al.; Endocrinology 145, 2747 (2004), Abstract;
Inhibition of metalloproteinase cleavage enhances the cytotoxicity of Fas ligand: P.G. Knox, et al.; J. Immunol. 170, 677 (2003), Abstract; Full Text
NF-kappa B is required for surface Ig-induced Fas resistance in B cells: B.R. Schram & T.L. Rothstein; J. Immunol. 170, 3118 (2003), Abstract; Full Text
Theileria parva-Transformed T Cells Show Enhanced Resistance to Fas/Fas Ligand-Induced Apoptosis : P. Küenzi, et al.; J. Immunol. 171, 1224 (2003), Application(s): Death induction of Theileria parva-infected TpM(803) T cells, Abstract; Full Text
TNF-related apoptosis-inducing ligand (TRAIL) frequently induces apoptosis in Philadelphia chromosome-positive leukemia cells: K. Uno, et al.; Blood 101, 3658 (2003), Application(s): Death induction of 12 Ph1-positive leukemia cell lines, Abstract; Full Text
Regulation of Fas (CD95)-induced apoptosis by nuclear factor-kappa B and tumor necrosis factor-alpha in macrophages: B. Lu, et al.; Am. J. Physiol. Cell Physiol. 283, C831 (2002), Application(s): Death induction of RAW 264.7 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
Tumor-cell resistance to death receptor-induced apoptosis through mutational inactivation of the proapoptotic Bcl-2 homolog Bax: H. LeBlanc, et al.; Nat. Med. 8, 274 (2002), Abstract;
Fas ligand (CD95L) protects neurons against perforin-mediated T lymphocyte cytotoxicity: I. Medana, et al.; J. Immunol. 167, 674 (2001), Abstract; Full Text
Fist/Hipk3: A FAS/Fadd-Interacting Serine/Threonine Kinase That Induces Fadd Phosphorylation and Inhibits FAS-Mediated Jun Nh2-Terminal Kinase Activation: V. Rochat-Steiner, et al.; J. Exp. Med. 192, 1165 (2000), Application(s): Death induction of Jurkat and JNK activation of 293T cells, Abstract; Full Text
General Literature References
Death receptors: signaling and modulation: A. Ashkenazi & V.M. Dixit; Science 281, 1305 (1998), Abstract;
Apoptosis by death factor: S. Nagata; Cell 88, 355 (1997), Abstract;
Melanoma cell expression of Fas(Apo-1/CD95) ligand: implications for tumor immune escape: M. Hahne, et al.; Science 274, 1363 (1996), Abstract;
A role for CD95 ligand in preventing graft rejection: D. Bellgrau, et al.; Nature 377, 630 (1995), Abstract;
Mature T cells of autoimmune lpr/lpr mice have a defect in antigen-stimulated suicide: J.H. Russel, et al.; PNAS 90, 4409 (1993), Abstract;
High activity, high purity CD40L protein for co-stimulatory activation of an immune response
Produced in CHO cells. The extracellular domain of human CD40L (CD154) (aa 116-261) is fused at the N-terminus to mouse ACRP30headless (aa 18-111) and a FLAG®-tag., ≥90% (SDS-PAGE) | Print as PDF
High activity, high purity CD40L protein for co-stimulatory activation of an immune response
Produced in CHO cells. The extracellular domain of mouse CD40L (CD154) (aa 115-260) is fused at the N-terminus to mouse ACRP30headless (aa 18-111) and a FLAG®-tag., ≥95% (SDS-PAGE) | Print as PDF
Enhanced ligand that utilizes the proprietary Killer™ linker peptide that promotes stability.
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., ≥99% (SDS-PAGE, MS analysis) | Print as PDF
Enhanced ligand with improved stability providing significantly enhanced immune activation.
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., ≥98% (SDS-PAGE). MS analysis, <1% impurity (mainly Hsp70 protein from E. coli) | Print as PDF
