hh Rat Fas Ligand/TNFSF6 Antibody AF1858: R&D Systems

Rat Fas Ligand/TNFSF6 Antibody

  
  • Species Reactivity
    Rat
  • Specificity
    Detects rat Fas Ligand/TNFSF6 in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 20% cross-reactivity with recombinant mouse Fas Ligand is observed and 5% cross-reactivity with recombinant human Fas Ligand is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant rat Fas Ligand/TNFSF6
    Leu104-Leu278
    Accession # P36940
  • 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 Rat Fas Ligand/TNFSF6 (Catalog # 1614-FL)
  • Immunocytochemistry
    5-15 µg/mL
    See below
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
Immunocytochemistry
Fas Ligand/TNFSF6 in Rat Splenocytes. Fas Ligand/TNFSF6 was detected in immersion fixed rat splenocytes using Goat Anti-Rat Fas Ligand/TNFSF6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1858) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
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 rat Fas Ligand consists of a 179 amino acid (aa) extracellular domain (ECD), a 22 aa transmembrane segment, and a 79 aa cytoplasmic domain. Within the ECD, rat Fas Ligand shares 78% and 93% aa sequence identity with human and mouse Fas Ligand, respectively. Fas Ligand is expressed as a nondisulfide-linked homotrimer on the surface of 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. 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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