Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Immunohistochemistry, Western Blot, Neutralization, Flow Cytometry

Cited:

Western Blot, Neutralization, Flow Cytometry, Immunocytochemistry, Bioassay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all Fas Ligand/TNFSF6 Primary Antibodies »

Product Specifications

Immunogen

Chinese hamster ovary cell line CHO-derived recombinant human Fas Ligand/TNFSF6
Pro134-Leu281
Accession # Q53ZZ1

Specificity

Detects human Fas Ligand in direct ELISAs and Western blots. In direct ELISAs, approximately 20% cross-reactivity with recombinant mouse Fas Ligand is observed, approximately 7% cross-reactivity with recombinant rat Fas Ligand and recombinant human (rh) BAFF, and less than 1% cross-reactivity with rhTRAIL, rhTNF-alpha, rhGITR Ligand, and rhAPRIL is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human Fas Ligand/TNFSF6 Antibody

Detection of Fas Ligand/TNFSF6 antibody in HEK293 Human Cell Line Transfected with Human Fas Ligand/TNFSF6 and eGFP antibody by Flow Cytometry.

Detection of Fas Ligand/TNFSF6 in HEK293 Human Cell Line Transfected with Human Fas Ligand/TNFSF6 and eGFP by Flow Cytometry.

HEK293 human embryonic kidney cell line transfected with (A) Fas Ligand/TNFSF6 or (B) irrelevant protein, and eGFP were stained with Goat Anti-Human Fas Ligand/TNFSF6 Affinity Purified Polyclonal Antibody (Catalog # AF126) followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). Quadrant markers were set based Goat IgG Control Antibody staining (Catalog # AB-108-C, data not shown). View our protocol for Staining Membrane-associated Proteins.
Fas Ligand/TNFSF6 antibody in Human Melanoma by Immunohistochemistry (IHC-P).

Fas Ligand/TNFSF6 in Human Melanoma.

Fas Ligand/TNFSF6 was detected in immersion fixed paraffin-embedded sections of human melanoma tissue using Goat Anti-Human Fas Ligand/TNFSF6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF126) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of immersion fixed paraffin-embedded Tissue Sections.

Fas Ligand/TNFSF6 in Human Thymus.

Fas Ligand/TNFSF6 was detected in immersion fixed paraffin-embedded sections of human thymus using Goat Anti-Human Fas Ligand/TNFSF6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF126) at 15 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013)). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Apoptosis Induced by Fas Ligand/TNFSF6 and Neutralization by Human Fas Ligand/TNFSF6 Antibody.

Apoptosis Induced by Fas Ligand/TNFSF6 and Neutralization by Human Fas Ligand/TNFSF6 Antibody.

In the presence of a cross-linking antibody, Mouse polyHistidine Monoclonal Antibody (10 µg/mL, Catalog # MAB050), Recombinant Human Fas Ligand/TNFSF6 (Catalog # 126-FL) induces apoptosis in the Jurkat human acute T cell leukemia cell line in a dose-dependent manner (orange line). Apoptosis elicited by Recombinant Human Fas Ligand/TNFSF6 (10 ng/mL) is neutralized (green line) by increasing concentrations of Human Fas Ligand/TNFSF6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF126). The ND50 is typically 0.012-0.072 µg/mL.
Detection of Human Fas Ligand/TNFSF6 by Western Blot

Detection of Human Fas Ligand/TNFSF6 by Western Blot

Different effects of cell release by trypsin and NECDS on subsequent initiation of anoikis and on antibody accessibility to the cell surface anoikis-initiating molecules in MUC1-positive and -negative cells. (a) HCA1.7+/− cells were released by NECDS or trypsin and cultured in suspension for 24 h before the cell-associated caspase-3/-7 activities were assessed. The data are presented as mean±S.E.M. of triplicate determinations of two independent experiments. (b–d) Representative flow cytometry plots show antibody access to cell surface MUC1 (b), E-cadherin, Integrin beta 1, CD44 (c, d) and recombinant Fas-L access to cell surface Fas (f) in HCA1.7+/− cells released by trypsin or NECDS. Note, an additional integrin beta 1 peak (arrowed) is seen in HCA1.7− cells released by NECDS in comparison to those released by trypsin. (e) Immunoblotting of cell lysates shows total cellular expression of CD44, E-cadherin, integrin beta 1, Fas, Fas-L and tubulin in HCA1.7+/− cells Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25275599), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human Fas Ligand/TNFSF6 Antibody

Application
Recommended Usage

Flow Cytometry

0.25 µg/mL
Sample: HEK293 Human Cell Line Transfected with Human Fas Ligand/TNFSF6 and eGFP 

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human melanoma tissue and human thymus

Western Blot

0.1 µg/mL
Sample: Recombinant Human Fas Ligand/TNFSF6 (Catalog # 126-FL)

Neutralization

Measured by its ability to neutralize Fas Ligand/TNFSF6-induced apoptosis in the Jurkat human acute T cell leukemia cell line. Cifone, M.G. et al. (1994) J. Exp. Med. 180:1547. The Neutralization Dose (ND50) is typically 0.012-0.072 μg/mL in the presence of 10 ng/mL Recombinant Human Fas Ligand/TNFSF6 and 10 µg/mL of a cross-linking antibody, Mouse polyHistidine Monoclonal Antibody.

Reviewed Applications

Read 1 review rated 5 using AF126 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store 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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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 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).

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. USA 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.

Alternate Names

CD178, CD95L, FASLG, TNFSF6

Entrez Gene IDs

356 (Human); 14103 (Mouse); 25385 (Rat)

Gene Symbol

FASLG

UniProt

Q53ZZ1

Additional Fas Ligand/TNFSF6 Products

Product Documents for Human Fas Ligand/TNFSF6 Antibody

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Product Specific Notices for Human Fas Ligand/TNFSF6 Antibody

For research use only

Citations for Human Fas Ligand/TNFSF6 Antibody

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  • Human Fas Ligand/TNFSF6 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Cancer Cells
    Species: Human
    Verified Customer | Posted 10/17/2018
    Human Fas Ligand/TNFSF6 Antibody AF126

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