Fas Ligand/TNFSF6 Antibody (154911) [Unconjugated]

FasL was detected in immersion fixed paraffin-embedded sections of human Tonsil using Mouse Anti-Human FasL Monoclonal Antibody (Catalog # MAB0951) at 3ug/mL at 37 ° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH). Tissue was stained using the Alexa Fluor™ 647 Goat anti-Mouse IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647MS) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the membrane. Protocol available in COMET™ Panel Builder.​​

Fas Ligand/TNFSF6 was detected in immersion fixed paraffin-embedded sections of human thymus (Hassall's corpuscle) using Mouse Anti-Human Fas Ligand/TNFSF6 Monoclonal Antibody (Catalog # MAB0951) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Western Blot shows lysates of HMCB human bowes melanoma cell line, NK‑92 human natural killer lymphoma cell line, SW480 human colorectal adenocarcinoma cell line, human placenta and human thymus. PVDF membrane was probed with 2 µg/ml of Mouse Anti-Human Fas Ligand/TNFSF6 Monoclonal Antibody (Catalog # MAB0951) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Fas Ligand/TNFSF6 at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

• Reactivity: Hu
• Applications: WB, IHC, mIF
• Clone: 154911
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

Fas Ligand/TNFSF6 Antibody (154911) [Unconjugated] Summary

• Immunogen: Chinese Hamster Ovary cell line, CHO-derived human Fas Ligand/TNFSF6
  Pro134-Leu281
  Accession # P48023
• Specificity: Detects human Fas Ligand/TNFSF6 in direct ELISAs.
• Source: N/A
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-25 ug/mL
  • Multiplex Immunofluorescence 3 ug/mL
  • Western Blot 2 ug/mL

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.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
  *Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

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

Alternate Names for Fas Ligand/TNFSF6 Antibody (154911) [Unconjugated]

• 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

Background

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).

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Uniprot:
  • P48023()