Human Fas Ligand/TNFSF6 Antibody

(3 citations)   
  • 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 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)
  • 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 2 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.
Data Examples
Immunohistochemistry
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
Fas Ligand/TNFSF6 in Human Melanoma. Fas Ligand/TNFSF6 was detected in immersion fixed paraffin-embedded sections of human melanoma 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). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
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 (2 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 0.12-0.072 μg/mL.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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
Related Research Areas
Citations:

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.

3 Citations: Showing 1 - 3
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Species
Applications
Sample Type
  1. 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;7(5):e35399.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  2. 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;7(6):e39576.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  3. 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;116(4):923-4.
    Species: Human
    Sample Type: Serum
    Application: Neut
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