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Human Fas Ligand/TNFSF6 Antibody

Catalog #: AF126
7 Citations 1 Review Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
AF126
AF126-SP
Detection of Fas Ligand/TNFSF6 in HEK293 Human Cell Line Transfected with Human Fas Ligand/TNFSF6 and eGFP by Flow Cytometry.
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Citations (7)
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Reviews (1)
Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human Fas Ligand/TNFSF6 Antibody Summary

Species Reactivity
Human
Specificity
Detects human Fas Ligand in direct ELISAs and Western blots. In direct ELISAs, approximately 20% cross-reactivity with recombinant mouse Fas Ligand is observed, approximately 7% cross-reactivity with recombinant rat Fas Ligand and recombinant human (rh) BAFF, and less than 1% cross-reactivity with rhTRAIL, rhTNF-alpha, rhGITR Ligand, and rhAPRIL is observed.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
Chinese hamster ovary cell line CHO-derived recombinant human Fas Ligand/TNFSF6
Pro134-Leu281
Accession # Q53ZZ1
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.1 µg/mL
Recombinant Human Fas Ligand/TNFSF6 (Catalog # 126-FL)
Flow Cytometry
0.25 µg/mL
See below
Immunohistochemistry
5-15 µg/mL
See below
Neutralization
Measured by its ability to neutralize Fas Ligand/TNFSF6-induced apoptosis in the Jurkat human acute T cell leukemia cell line. Cifone, M.G. et al. (1994) J. Exp. Med. 180:1547. The Neutralization Dose (ND50) is typically 0.012-0.072 μg/mL in the presence of 10 ng/mL Recombinant Human Fas Ligand/TNFSF6 and 10 µg/mL of a cross-linking antibody, Mouse polyHistidine Monoclonal Antibody.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Flow Cytometry Detection of Fas Ligand/TNFSF6 antibody in HEK293 Human Cell Line Transfected with Human Fas Ligand/TNFSF6 and eGFP antibody by Flow Cytometry. View Larger

Detection of Fas Ligand/TNFSF6 in HEK293 Human Cell Line Transfected with Human Fas Ligand/TNFSF6 and eGFP by Flow Cytometry. HEK293 human embryonic kidney cell line transfected with (A) Fas Ligand/TNFSF6 or (B) irrelevant protein, and eGFP were stained with Goat Anti-Human Fas Ligand/TNFSF6 Affinity Purified Polyclonal Antibody (Catalog # AF126) followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). Quadrant markers were set based Goat IgG Control Antibody staining (Catalog # AB-108-C, data not shown). View our protocol for Staining Membrane-associated Proteins.

Immunohistochemistry Fas Ligand/TNFSF6 antibody in Human Melanoma by Immunohistochemistry (IHC-P). View Larger

Fas Ligand/TNFSF6 in Human Melanoma. Fas Ligand/TNFSF6 was detected in immersion fixed paraffin-embedded sections of human melanoma tissue using Goat Anti-Human Fas Ligand/TNFSF6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF126) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of immersion fixed paraffin-embedded Tissue Sections.

Immunohistochemistry View Larger

Fas Ligand/TNFSF6 in Human Thymus. Fas Ligand/TNFSF6 was detected in immersion fixed paraffin-embedded sections of human thymus using Goat Anti-Human Fas Ligand/TNFSF6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF126) at 15 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013)). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Neutralization Apoptosis Induced by Fas Ligand/TNFSF6 and Neutralization by Human Fas Ligand/TNFSF6 Antibody. View Larger

Apoptosis Induced by Fas Ligand/TNFSF6 and Neutralization by Human Fas Ligand/TNFSF6 Antibody. In the presence of a cross-linking antibody, Mouse polyHistidine Monoclonal Antibody (10 µg/mL, Catalog # MAB050), Recombinant Human Fas Ligand/TNFSF6 (Catalog # 126-FL) induces apoptosis in the Jurkat human acute T cell leukemia cell line in a dose-dependent manner (orange line). Apoptosis elicited by Recombinant Human Fas Ligand/TNFSF6 (10 ng/mL) is neutralized (green line) by increasing concentrations of Human Fas Ligand/TNFSF6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF126). The ND50 is typically 0.012-0.072 µg/mL.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Fas Ligand/TNFSF6

Fas Ligand (FasL), also known as CD178, CD95L, or TNFSF6, is a 40 kDa type II transmembrane member of the TNF superfamily of proteins. Its ability to induce apoptosis in target cells plays an important role in the development, homeostasis, and function of the immune system (1). Mature human Fas Ligand consists of a 179 amino acid (aa) extracellular domain (ECD), a 22 aa transmembrane segment, and a 80 aa cytoplasmic domain (2). Within the ECD, human Fas Ligand shares 81% and 78% aa sequence identity with mouse and rat Fas Ligand, respectively. Both mouse and human Fas Ligand are active on mouse and human cells (2, 3). Fas Ligand is expressed on the cell surface as a nondisulfide-linked homotrimer on activated CD4+ Th1 cells, CD8+ cytotoxic T cells, and NK cells (1). Fas Ligand binding to Fas/CD95 on an adjacent cell triggers apoptosis in the Fas‑expressing cell (2, 4). Fas Ligand also binds DcR3 which is a soluble decoy receptor that interferes with Fas Ligand-induced apoptosis (5). Fas Ligand can be released from the cell surface by metalloproteinases as a 26 kDa soluble molecule which remains trimeric (6, 7). Shed Fas Ligand retains the ability to bind Fas, although its ability to trigger apoptosis is dramatically reduced (6, 7). In the absence of TGF‑ beta, however, Fas Ligand/Fas interactions instead promote neutrophil-mediated inflammatory responses (3, 8). Fas Ligand itself transmits reverse signals that costimulate the proliferation of freshly antigen-stimulated T cells (9). Fas Ligand-induced apoptosis plays a central role in the development of immune tolerance and the maintance of immune privileged sites (10). This function is exploited by tumor cells which evade immune surveillance by upregulating Fas Ligand to kill tumor infiltrating lymphocytes (8, 11). In gld mice, a Fas Ligand point mutation is the cause of severe lymphoproliferation and systemic autoimmunity (12, 13).

References
  1. Lettau, M. et al. (2008) Curr. Med. Chem. 15:1684.
  2. Takahashi, T. et al. (1994) Int. Immunol. 6:1567.
  3. Seino, K-I. et al. (1998) J. Immunol. 161:4484.
  4. Suda, T. et al. (1993) Cell 75:1169.
  5. Pitti, R.M. et al. (1998) Nature 396:699.
  6. Schneider, P. et al. (1998) J. Exp. Med. 187:1205.
  7. Tanaka, M. et al. (1998) Nature Med. 4:31.
  8. Chen, J-J. et al. (1998) Science 282:1714.
  9. Suzuki, I. and P.J. Fink (2000) Proc. Natl. Acad. Sci. USA 97:1707.
  10. Ferguson, T.A. and T.S. Griffith (2006) Immunol. Rev. 213:228.
  11. Ryan, A.E. et al. (2005) Cancer Res. 65:9817.
  12. Takahashi, T. et al. (1994) Cell 76:969.
  13. Lynch, D.H. et al. (1994) Immunity 1:131.
Entrez Gene IDs
356 (Human); 14103 (Mouse); 25385 (Rat)
Alternate Names
apoptosis (APO-1) antigen ligand 1; Apoptosis antigen ligand; APT1LG1CD95L; APTL; CD178 antigen; CD178; CD95L; CD95-L; Fas antigen ligand; Fas ligand (TNF superfamily, member 6); Fas Ligand; FASLCD95 ligand; FASLG; TNFSF6; TNFSF6FasL; tumor necrosis factor (ligand) superfamily, member 6; tumor necrosis factor ligand superfamily member 6

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Citations for Human Fas Ligand/TNFSF6 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

7 Citations: Showing 1 - 7
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  1. MUC1 O-glycosylation contributes to anoikis resistance in epithelial cancer cells
    Authors: Tushar Piyush, Jonathan M Rhodes, Lu-Gang Yu
    Cell Death Discovery
  2. Tissue Inhibitor of Metalloproteinase-3 (TIMP-3) induces FAS dependent apoptosis in human vascular smooth muscle cells
    Authors: WR English, H Ireland-Ze, AH Baker, TD Littlewood, MR Bennett, G Murphy
    PLoS ONE, 2018-04-04;13(4):e0195116.
    Species: Human
    Sample Types: Cell Lysates, Whole Cells
    Applications: Flow Cytometry, Neutralization, Western Blot
  3. Human cytomegalovirus infection elicits new decidual natural killer cell effector functions.
    Authors: Siewiera J, El Costa H, Tabiasco J, Berrebi A, Cartron G, Le Bouteiller P, Jabrane-Ferrat N
    PLoS Pathog, 2013-04-04;9(4):e1003257.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  4. Activated Human CD4+CD45RO+ Memory T-Cells Indirectly Inhibit NLRP3 Inflammasome Activation through Downregulation of P2X7R Signalling.
    Authors: Beynon V, Quintana FJ, Weiner HL
    PLoS ONE, 2012-06-29;7(6):e39576.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  5. Thymosin beta10 expression driven by the human TERT promoter induces ovarian cancer-specific apoptosis through ROS production.
    Authors: Kim YC, Kim BG, Lee JH
    PLoS ONE, 2012-05-18;7(5):e35399.
    Species: Human
    Sample Types: Cell Lysates, Whole Cells
    Applications: ICC, Western Blot
  6. Rapid resolution of toxic epidermal necrolysis with anti-TNF-alpha treatment.
    Authors: Hunger RE, Hunziker T, Buettiker U, Braathen LR, Yawalkar N
    J. Allergy Clin. Immunol., 2005-08-19;116(4):923-4.
    Species: Human
    Sample Types: Serum
    Applications: Neutralization
  7. MUC1 extracellular domain confers resistance of epithelial cancer cells to anoikis
    Authors: Q Zhao, T Piyush, C Chen, M A Hollingsworth, J Hilkens, J M Rhodes et al.
    Cell Death & Disease

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Human Fas Ligand/TNFSF6 Antibody
By Anonymous on 10/17/2018
Application: WB Sample Tested: Cancer Cells Species: Human
Western Blot Human Fas Ligand/TNFSF6 Antibody AF126