Recombinant Mouse Fas/TNFRSF6/CD95 Fc Chimera Protein

• Applications: Bioactivity

Recombinant Mouse Fas/TNFRSF6/CD95 Fc Chimera Protein Summary

• Details of Functionality: Measured by its ability to inhibit Fas Ligand-induced apoptosis of Jurkat human acute T cell leukemia cells. The ED₅₀ for this effect is 0.03‑0.1 µg/mL in the presence of 20 ng/mL recombinant human Fas-ligand.
• Source: Spodoptera frugiperda, Sf 21 (baculovirus)-derived mouse Fas/TNFRSF6/CD95 protein
  

  Mouse FAS (Gly14 - Arg169) Accession # P25446	DIEGRMD	Human IgG1 (Pro100 - Lys330)	6-His tag
  N-terminus			C-terminus

• Accession #: P25446
• N-terminal Sequence: Gly14
• Structure / Form: Disulfide-linked homodimer
• Protein/Peptide Type: Recombinant Proteins
• Gene: Fas
• Purity: >97%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 46 kDa (monomer).
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 55 kDa, reducing conditions

Packaging, Storage & Formulations

• 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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
• Purity: >97%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Reconstitution Instructions: Reconstitute at 50 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Mouse Fas/TNFRSF6/CD95 Fc Chimera Protein

• 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

Background

Fas, also known as APO‑1 or CD95, belongs to the death receptor subfamily of the TNF receptor superfamily and is designated TNFRSF6 (1 ‑ 3). Mouse Fas cDNA encodes 327 amino acids (aa) that include a 21 aa signal peptide, a 148 aa extracellular domain (ECD) that contains three cysteine‑ich TNFR repeats, a 17 aa transmembrane sequence, and a 141 aa cytoplasmic domain containing a death domain (DD), which is required for transducing apoptotic signals (4). Mature mouse Fas ECD shares 68% aa sequence identity with rat Fas, and 53 ‑ 55% with human, feline, bovine and porcine Fas. One potentially secreted 63 aa isoform diverges at aa 61 (5). The Fas ligand (FasL, TNFSF6) is a type II transmembrane protein of the TNF family that can be expressed on activated T‑lymphocytes, NK cells and cells in immune privileged sites, or shed in soluble form (2). Polymorphisms of FasL impart differing activity of Fas depending on mouse strain (6). Engagement of FAS induces oligomerization of preformed Fas trimers (1, 2). The activated receptor recruits the adaptor molecule FADD to form the Death‑Inducing Signaling Complex (DISC). Upon activation, caspases in the DISC initiate the apoptotic signaling cascade (7). Fas is prominent in epithelial cells, hepatocytes, activated mature lymphocytes, virus‑transformed lymphocytes and tumor cells. It is an essential mediator in the activation‑induced death of T lymphocytes that terminates the immune reaction (1, 2, 8). In immune‑privileged tissues, infiltrating Fas‑bearing lymphocytes and inflammatory cells are killed by FasL engagement (9). Both humans and mice with genetic defects in Fas accumulate abnormal lymphocytes and develop systemic autoimmunity (1 ‑ 3). The Fas pathway also appears to cross‑communicate with the BIM (mitochondrial/intrinsic) apoptosis pathway (1). 

1. Bouillet, P. and L.A. O’Reilly (2009) Nat. Rev. Immunol. 9:514.
2. Strasser, A. et al. (2009) Immunity 30:180.
3. Ashkenazi, A. and V. Dixit (1999) Curr. Opin. Cell Biol. 11:255.
4. SwissProt accession P25446.
5. NCBI accession NP_001140180.
6. Kayagaki, N. et al. (1997) Proc. Natl. Acad. Sci. USA 94:3914.
7. Thorburn, A. (2003) Cellular Signaling 16:139.
8. Barreiro, R. et al. (2004) J. Immunol. 173:1519.
9. Ferguson, T.A. and T.S Griffith (2006) Immunol. Rev. 213:228.

Publications for Fas/TNFRSF6/CD95 (435-FA)(5)

Publications using 435-FA

• M Kamiya, F Mizoguchi, K Kawahata, D Wang, M Nishibori, J Day, C Louis, IP Wicks, H Kohsaka, S Yasuda Targeting necroptosis in muscle fibers ameliorates inflammatory myopathies Nature Communications, 2022-01-10;13(1):166. 2022-01-10 [PMID: 35013338] (Bioassay, Mouse)
• IK Choi, Z Wang, Q Ke, M Hong, Y Qian, X Zhao, Y Liu, HJ Kim, J Ritz, H Cantor, K Rajewsky, KW Wucherpfen, B Zhang Signaling by the Epstein-Barr virus LMP1 protein induces potent cytotoxic CD4+ and CD8+ T cell responses Proc. Natl. Acad. Sci. U.S.A., 2018-01-08;0(0):. 2018-01-08 [PMID: 29311309] (Bioassay, Mouse)
• Deegan S, Saveljeva S, Logue S, Pakos-Zebrucka K, Gupta S, Vandenabeele P, Bertrand M, Samali A Deficiency in the mitochondrial apoptotic pathway reveals the toxic potential of autophagy under ER stress conditions. Autophagy, 2014-01-01;10(11):1921-36. 2014-01-01 [PMID: 25470234] (Bioassay, Mouse)
• Wetzel M, Li L, Harms KM, Roitbak T, Ventura PB, Rosenberg GA, Khokha R, Cunningham LA Tissue inhibitor of metalloproteinases-3 facilitates Fas-mediated neuronal cell death following mild ischemia. Cell Death Differ., 2007-10-26;15(1):143-51. 2007-10-26 [PMID: 17962815] (Bioassay, Mouse)
• Bheeshmachar G, Purushotaman D, Sade H, Gunasekharan V, Rangarajan A, Sarin A Evidence for a role for notch signaling in the cytokine-dependent survival of activated T cells. J. Immunol., 2006-10-15;177(8):5041-50. 2006-10-15 [PMID: 17015687] (Bioassay, Mouse)

Bioinformatics

• Gene Symbol: Fas
• Uniprot:
  • P25446()