Feline Fas/TNFRSF6/CD95 Biotinylated Antibody

Catalog #: BAF2267 Datasheet
Catalog # Availability Size / Price Qty
BAF2267
Detection of Fas/TNFRSF6/ CD95 in Feline PBMCs treated with PMA and Ca2+ionomycin by Flow Cytometry.
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Feline Fas/TNFRSF6/CD95 Biotinylated Antibody Summary

Species Reactivity
Feline
Specificity
Detects feline Fas/TNFRSF6/CD95 in Western blots. In Western blots, less than 5% cross-reactivity with recombinant human Fas and recombinant mouse 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 BSA as a carrier protein.
Label
Biotin

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
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.

Scientific Data

Flow Cytometry Detection of Fas/TNFRSF6/ CD95 antibody in Feline PBMCs treated with PMA and Ca<sup>2+</sup>ionomycin antibody by Flow Cytometry. View Larger

Detection of Fas/TNFRSF6/ CD95 in Feline PBMCs treated with PMA and Ca2+ionomycin by Flow Cytometry. Feline peripheral blood mononuclear cells were treated with PMA and Ca2+ionomycin, then stained with Goat Anti-Feline Fas/TNFRSF6/ CD95 Biotinylated Antigen Affinity-purified Polyclonal Antibody (Catalog # BAF2267, filled histogram) or isotype control antibody (Catalog # BAF108, open histogram), followed by Streptavidin-Phycoerythrin (Catalog # F0040).

Immunocytochemistry Fas/TNFRSF6/CD95 antibody in Feline PBMCs by Immunocytochemistry (ICC). View Larger

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 Biotinylated Antigen Affinity-purified Polyclonal Antibody (Catalog # BAF2267) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Streptavidin (red; Catalog # NL999) 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.
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Shipping
The product is shipped at ambient temperature. Upon receipt, store it 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/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); 102140989 (Cynomolgus Monkey); 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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