Feline Fas/TNFRSF6/CD95 Antibody

  
  • Species Reactivity
    Feline
  • Specificity
    Detects feline Fas/TNFRSF6/CD95 in direct ELISAs and Western blots. In direct ELISAs and Western blots, less than 1% cross-reactivity with recombinant human Fas is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant feline Fas/TNFRSF6/CD95
    Ala25-Lys172
    Accession # NP_001009314
  • 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.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    Recombinant Feline Fas/TNFRSF6/CD95 Fc Chimera (Catalog # 2267-FA)
  • Flow Cytometry
    2.5 µg/106 cells
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • 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
Detection of Fas/TNFRSF6/
CD95 in Feline PBMCs by Flow Cytometry.
Feline peripheral blood mononuclear cells were stained with Feline Fas/TNFRSF6/CD95 Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF2267, filled histogram) or isotype control antibody (Catalog # AB‑108‑C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107).
Immunocytochemistry
Fas/TNFRSF6/CD95 in Feline PBMCs. Fas/TNFRSF6/CD95 was detected in immersion fixed feline peripheral blood mononuclear cells (PBMCs) using Goat Anti-Feline Fas/TNFRSF6/CD95 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2267) 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 the plasma membrane. 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/TNFRSF6/CD95

Feline Fas (fibroblast associated; also named CD95 and APO-1) is a 45 kDa type I transmembrane (TM) glycoprotein that is a member of the TNF receptor superfamily (1-3). The family contains about 30 members, and is characterized by the presence of at least one cysteine-rich domain that contains multiple intrachain disulfide bonds. In general, the superfamily is divided into cytoplasmic death domain (DD) containing, and non-DD containing receptors (3). Feline Fas is synthesized as a 314 amino acid (aa) precursor that contains a 24 aa signal sequence, a 148 aa extracellular region, a 16 aa TM segment, and a 126 aa cytoplasmic tail (4). The extracellular region contains four potential N-linked glycosylation sites plus two distinct cysteine-rich domains of approximately 40 aa each; the cytoplasmic tail shows a 45 aa DD. The extracellular region of feline Fas shares 68%, 65%, 53%, and 58% aa sequence identity to porcine, human, mouse, and rat Fas, respectively. There are five alternate splice forms of feline Fas, which vary from 132 aa to 209 aa in length. All utilize exons 1-3 (aa 1-111) and all are missing the transmembrane segment of the full length form (5). Circulating Fas is reported to be both a dimer and trimer at low ng/mL concentrations. The ligand for Fas is FasL, and Fas ligation activates both the MEK cascade and FADD/caspase-8 pathway (7).

  • References:
    1. Locksley, R.M. et al. (2001) Cell 104:487.
    2. Gaur, U. and B.B. Aggarwal (2003) Biochem. Pharmacol. 66:1403.
    3. Collette, Y. et al. (2003) Trends Immunol. 24:387.
    4. Mizuno, T. et al. (1998) Vet. Immunol. Immunopathol. 65:161.
    5. Mizuno, T. et al. (2004) Eur. J. Immunogenet. 31:159.
    6. Knipping, E. et al. (1995) Blood 85:1562.
    7. Baker, S.J. and E.P. Reddy (1998) Oncogene 17:3261.
  • Long Name:
    Fibroblast-associated
  • Entrez Gene IDs:
    355 (Human); 14102 (Mouse); 246097 (Rat); 493881 (Feline)
  • Alternate Names:
    Apo-1 antigen; Apo-1; apoptosis antigen 1; Apoptosis-mediating surface antigen FAS; APT1; APT1FASTM; CD95 antigen; CD95; CD95ALPS1A; Fas (TNF receptor superfamily, member 6); Fas AMA; Fas antigen; Fas; FAS1; FASLG receptor; TNFRSF6; TNFRSF6member 6; tumor necrosis factor receptor superfamily member 6
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