Recombinant Human Fas Ligand/TNFSF6 Protein

Catalog # 126-FL | R&D Systems, Inc. a Bio-Techne Brand
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Carrier Free
126-FL-010/CF
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With Carrier
126-FL-010
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Key Product Details

Accession #

Q53ZZ1

Source

CHO

Conjugate

Unconjugated

Applications

Bioactivity

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human Fas Ligand/TNFSF6 protein
Pro134-Leu281, with an N-terminal Met and 6-His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Met

Predicted Molecular Mass

18 kDa

SDS-PAGE

19 - 32 kDa, reducing conditions

Activity

Measured by its ability to induce apoptosis of Jurkat human acute T cell leukemia cells.
The ED50 for this effect is 0.3‑1.5 ng/mL in the presence of 10 µg/mL of a cross-linking antibody Mouse Anti-polyHistidine Monoclonal Antibody (Catalog # MAB050).

Formulation, Preparation and Storage

Carrier Free
What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

Carrier: 126-FL
Formulation Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
Reconstitution Reconstitute at 100 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin.
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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
Carrier Free: 126-FL/CF
Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution Reconstitute at 100 μg/mL in sterile PBS.
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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

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. 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

Product Documents for Recombinant Human Fas Ligand/TNFSF6 Protein

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS 126-FL (with carrier)
Download SDS 126-FL/CF (carrier free)

Product Specific Notices for Recombinant Human Fas Ligand/TNFSF6 Protein

For research use only

Citations for Recombinant Human Fas Ligand/TNFSF6 Protein (20)

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  • Recombinant Human Fas Ligand/TNFSF6 Protein
    Name: Giovanni Manzi
    Verified Customer | Posted 12/26/2022
    ***Bio-Techne Response: Thank you for reviewing our product. We are sorry to hear that this product did not perform as expected. We have been in touch with the customer to resolve this issue according to our Product Guarantee and to the customer’s satisfaction.*** ***The customer worked with our Technical Support team and had success detecting the complexed form of Complement Component C9 in the complement membrane attack complex (C5b-9) while using a polyclonal antibody for Human Complement Component C9 (AF8126). We have not tested complement membrane attack complex (C5b-9) detection with MAB8126 in-house.*** We used the ligand for a pulldown assay and unfortunately one of the major contaminating protein is the same size as our protein, so we can not analyse our experiment. It was an expensive experiment to run for a small amount of supposedly pure ligand.
    Recombinant Human Fas Ligand/TNFSF6 Protein 126-FL

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FAQs for Recombinant Human Fas Ligand/TNFSF6 Protein

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  • Q: This protein datasheet indicates I need to use a cross-linking antibody, Catalog # MAB050, for biological activity. What is this antibody and is it really necessary?

    A: The antibody is directed against a 6x histidine repeat and is recommended for use as a cross-linker of proteins with 6x his-tag. Crosslinking is often used for proteins that require receptor trimerization and can result greater biological activity. R&D Systems Quality Control tests the performance of these proteins in the presence of the cross-linking antibody. Therefore, it is necessary to use this antibody when trying to achieve the same level of specific activity described in the datasheet.

  • Q: Is Recombinant Human Fas Ligand/TNFSF6 Protein (Catalog # 126-FL) a trimer in solution?

    A: In house, we observed this 126-FL protein mostly in monomeric form with some dimers/trimers present while in solution. This protein can be oligamerized in the presence of the cross-linking antibody, Mouse Anti-polyHistidine Monoclonal Antibody (Catalog # MAB050).

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