Human Fas Ligand/TNFSF6 Antibody (100419)

R&D Systems | Catalog # MAB126

R&D Systems
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Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

ELISA Capture (Matched Antibody Pair), Neutralization, Flow Cytometry

Cited:

Neutralization, Functional Assay

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 100419
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 # P48023

Specificity

Detects human Fas Ligand/TNFSF6 in ELISAs.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Endotoxin Level

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

Scientific Data Images for Human Fas Ligand/TNFSF6 Antibody (100419)

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 Mouse Anti-Human Fas Ligand/TNFSF6 Monoclonal Antibody (Catalog # MAB126) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). Quadrant markers were set based Mouse IgG2B Isotype Control Antibody staining (Catalog # MAB0041, data not shown). View our protocol for Staining Membrane-associated Proteins.
Cytotoxicity Induced by Fas Ligand/TNFSF6 and Neutralization by Human Fas Ligand/TNFSF6 Antibody.

Cytotoxicity 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 (Catalog # MAB050), Recombinant Human Fas Ligand/TNFSF6 (Catalog # 126-FL) induces cytotoxicity in the Jurkat human acute T cell leukemia cell line in a dose-dependent manner (orange line). Under these conditions, cytotoxicity elicited by Recombinant Human Fas Ligand/TNFSF6 (2 ng/mL) is neutralized (green line) by increasing concentrations of Human Fas Ligand/TNFSF6 Monoclonal Antibody (Catalog # MAB126). The ND50 is typically 1-5 ng/mL.
Human Fas Ligand / TNFSF6 ELISA Standard Curve

Human Fas Ligand / TNFSF6 ELISA Standard Curve

Recombinant Human Fas Ligand/TNFSF6 (Catalog # 126-FL) was serially diluted and captured by Mouse Anti-Human Fas Ligand/TNFSF6 Monoclonal Antibody (Catalog # MAB126) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human Fas Ligand/TNFSF6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF126) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)

Applications for Human Fas Ligand/TNFSF6 Antibody (100419)

Application
Recommended Usage

Flow Cytometry

0.25 µg/106 cells
Sample: HEK293 Human Cell Line Transfected with Human Fas Ligand/TNFSF6 and eGFP

Neutralization

Measured by its ability to neutralize Fas Ligand/TNFSF6-induced cytotoxicity in the Jurkat human acute T cell leukemia cell line. The Neutralization Dose (ND50) is typically 1-5 ng/mL in the presence of 2 ng/mL Recombinant Human Fas Ligand/TNFSF6 and 10 µg/mL of a cross-linking antibody, Mouse poly-Histidine Monoclonal Antibody.

Human Fas Ligand/TNFSF6 Sandwich Immunoassay

ELISA Capture (Matched Antibody Pair)
Recommended Concentration: 2-8 µg/mL
Use in combination with these reagents:
  • Detection Reagent: Human Fas Ligand/TNFSF6 Biotinylated Antibody (Catalog # BAF126)
  • Standard: Recombinant Human Fas Ligand/TNFSF6 Protein (Catalog # 126-FL)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 2 reviews rated 4.5 using MAB126 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 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

P48023

Additional Fas Ligand/TNFSF6 Products

Product Documents for Human Fas Ligand/TNFSF6 Antibody (100419)

Certificate of Analysis

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Product Specific Notices for Human Fas Ligand/TNFSF6 Antibody (100419)

For research use only

Citations for Human Fas Ligand/TNFSF6 Antibody (100419)

Customer Reviews for Human Fas Ligand/TNFSF6 Antibody (100419) (2)

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2 Customer Ratings
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  • Human Fas Ligand/TNFSF6 Antibody
    Name: Anonymous
    Application: Block/Neutralize
    Sample Tested: Human NKT cell, Human CAR-T cell and activated CAR-T cell
    Species: Human
    Verified Customer | Posted 07/17/2018
    the Fas/FasL pathways in CAR-T cell apoptosis is significant rescued upon blockade by combined Anti-Fas and Anti-FasL Antibody.
    Human Fas Ligand/TNFSF6 Antibody (100419) MAB126
  • Human Fas Ligand/TNFSF6 Antibody
    Name: Preethi Vijayaraj
    Application: Immunohistochemistry
    Sample Tested: Adult lung
    Species: Human
    Verified Customer | Posted 11/04/2016

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