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Mouse Fas Ligand/TNFSF6 Antibody

Catalog #: MAB5262
7 Citations 1 Review Datasheet / COA / SDS
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MAB5262
MAB5262-SP

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Cytotoxicity Induced by Fas Ligand/TNFSF6 and Neutralization by Mouse Fas Ligand/TNFSF6 Antibody.
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Mouse Fas Ligand/TNFSF6 Antibody Summary

Species Reactivity
Mouse
Specificity
Detects mouse Fas Ligand/TNFSF6 in ELISAs and Western blots. In Western blots, approximately 2% cross-reactivity with recombinant mouse (rm) TNF-alpha  is observed and no cross-reactivity with recombinant human (rh) APRIL, rhFas Ligand, rhGITR Ligand, rhLIGHT, rhTRAIL, rmTRANCE, and rhVEGI is observed.
Source
Monoclonal Rat IgG1 Clone # 101626
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Mouse myeloma cell line NS0-derived recombinant mouse Fas Ligand/TNFSF6
Pro132-Leu279
Accession # P41047
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.
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
Recombinant Mouse Fas Ligand/TNFSF6 (Catalog # 526-SA)

Mouse Fas Ligand/TNFSF6 Sandwich Immunoassay

Recommended Concentration
Reagent
ELISA Capture (Matched Antibody Pair)
2-8 µg/mL 

Use in combination with:

Detection Reagent: Mouse Fas Ligand/TNFSF6 Biotinylated Antibody (Catalog # BAF526)

Standard: Recombinant Mouse Fas Ligand/TNFSF6 Protein (Catalog # 526-SA)

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 5-20 µg/mL in the presence of 5 µg/mL Recombinant Mouse Fas Ligand/TNFSF6 and 10 µg/mL of a cross-linking antibody, Mouse polyHistidine Monoclonal Antibody.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Neutralization Cytotoxicity Induced by Fas Ligand/TNFSF6 and Neutralization by Mouse Fas Ligand/TNFSF6 Antibody. View Larger

Cytotoxicity Induced by Fas Ligand/TNFSF6 and Neutralization by Mouse Fas Ligand/TNFSF6 Antibody. In the presence of a cross-linking antibody, Mouse polyHisti-dine Monoclonal Antibody (10 µg/mL, Catalog # MAB050), Recom-binant Mouse Fas Ligand/ TNFSF6 (Catalog # 526-SA) 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 Mouse Fas Ligand/ TNFSF6 (5 µg/mL) is neutral-ized (green line) by increasing concentrations of Rat Anti-Mouse Fas Ligand/TNFSF6 Monoclonal Antibody (Catalog # MAB5262). The ND50 is typically 5-20 µg/mL.

Flow Cytometry Detection of Fas Ligand/TNFSF6 by Flow Cytometry View Larger

Detection of Fas Ligand/TNFSF6 by Flow Cytometry CD5+ B cells express higher levels of Fas ligand (FasL) than other splenic populations. Freshly isolated splenocytes from type II collagen/complete Freund's adjuvant (cII/CFA)-immunized, cII T-cell receptor transgenic mice were stained with anti-CD19, anti-CD5, and anti-FasL antibodies as described in Materials and methods. (a) Cells were initially gated as small nongranular lymphocytes by forward scatter and side scatter criteria (not shown) and then categorized by staining with anti-CD19 and anti-CD5 as T cells (CD5hi/CD19-), CD5+ B cells (CD5+/CD19+), B-2 cells (CD5-/CD19+), or double-negatives. (b) Comparison of FasL expression on CD5+ B cells from four mice, with representative disease severity scores shown in parentheses. Numbers in the box are mean fluorescence intensity (MFI) values of FasL staining on CD5+ B cells from the four mice. (c) Histograms and MFI values compared for lymphocyte subsets of two representative mice. (d) Average MFI values of FasL expression on gated splenic lymphocytes from mice with severe (n = 6), moderate (n = 6), and mild or no arthritis (n = 6). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/19706160), licensed under a CC-BY license. Not internally tested by R&D Systems.

Flow Cytometry Detection of Fas Ligand/TNFSF6 by Flow Cytometry View Larger

Detection of Fas Ligand/TNFSF6 by Flow Cytometry CD5+ B cells express higher levels of Fas ligand (FasL) than other splenic populations. Freshly isolated splenocytes from type II collagen/complete Freund's adjuvant (cII/CFA)-immunized, cII T-cell receptor transgenic mice were stained with anti-CD19, anti-CD5, and anti-FasL antibodies as described in Materials and methods. (a) Cells were initially gated as small nongranular lymphocytes by forward scatter and side scatter criteria (not shown) and then categorized by staining with anti-CD19 and anti-CD5 as T cells (CD5hi/CD19-), CD5+ B cells (CD5+/CD19+), B-2 cells (CD5-/CD19+), or double-negatives. (b) Comparison of FasL expression on CD5+ B cells from four mice, with representative disease severity scores shown in parentheses. Numbers in the box are mean fluorescence intensity (MFI) values of FasL staining on CD5+ B cells from the four mice. (c) Histograms and MFI values compared for lymphocyte subsets of two representative mice. (d) Average MFI values of FasL expression on gated splenic lymphocytes from mice with severe (n = 6), moderate (n = 6), and mild or no arthritis (n = 6). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/19706160), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
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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.

Background: Fas Ligand/TNFSF6

Fas Ligand (FasL) is a 40 kDa type II membrane protein belonging to the TNF family. In the new TNF super family nomenclature, FasL is referred to as TNFSF6. The specific receptor for FasL is Fas (CD95, Apo-1), a 45 kDa type I transmembrane protein that is a member of the TNF receptor family. FasL is predominantly expressed on activated T cells and NK cells, while Fas is expressed on various types of cells. The Fas/FasL system plays a crucial role in modulating immune response by inducing cell apoptosis to maintain homeostasis, self-tolerance of lymphocytes, and immune privilege. FasL was reported to be a potent chemoattractant for neutrophils, suggesting a novel proinflammatory function of this molecule. Like other members of the TNF family, the membrane-bound FasL can be cleaved by metalloproteinase to generate the soluble Fas ligand (sFasL) which is mainly a non-covalently linked homotrimer. It has been shown that the membrane-bound TNF‑ alpha and FasL are primary activators of their receptors. In contrast to soluble TNF‑ alpha which has potent cytotoxicity, sFasL is much less cytotoxic. Studies have shown that sFasL may competitively inhibit the killing effect of membrane FasL indicating that the cleaving of membrane FasL might be a mechanism to down‑regulate their activities.

References
  1. Suda, T. et al. (1993) Cell 75:1169.
  2. Kägi, D. et al. (1994) Science 265:528.
  3. Schneider, P. et al. (1998) J. Exp. Med. 187:1205.
  4. Seino, K. et al. (1998) J. Immunol. 161:4484.
Entrez Gene IDs
356 (Human); 14103 (Mouse); 25385 (Rat)
Alternate Names
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

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Citations for Mouse Fas Ligand/TNFSF6 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

7 Citations: Showing 1 - 7
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  1. Adenovirus-Mediated FasL Minigene Transfer Endows Transduced Cells with Killer Potential
    Authors: Madalina Dumitrescu, Violeta Georgeta Trusca, Lorand Savu, Ioana Georgeta Stancu, Attila Cristian Ratiu, Maya Simionescu et al.
    International Journal of Molecular Sciences
  2. FasL Modulates Expression of Mmp2 in Osteoblasts
    Authors: E Svandova, B Vesela, H Lesot, J Sadoine, A Poliard, E Matalova
    Front Physiol, 2018-09-19;9(0):1314.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Inhibition
  3. Glycolysis promotes caspase-3 activation in lipid rafts in T cells
    Authors: MA Secinaro, KA Fortner, O Dienz, A Logan, MP Murphy, V Anathy, JE Boyson, RC Budd
    Cell Death Dis, 2018-01-19;9(2):62.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  4. The molecular signature of murine T cell homeostatic proliferation reveals both inflammatory and immune inhibition patterns
    Authors: Karen A. Fortner, Jeffrey P. Bond, James W. Austin, Jeremy M. Boss, Ralph C. Budd
    Journal of Autoimmunity
  5. Response patterns of cytokines/chemokines in two murine strains after irradiation.
    Authors: Zhang M, Yin L, Zhang K, Sun W, Yang S, Zhang B, Salzman P, Wang W, Liu C, Vidyasagar S, Zhang L, Ju S, Okunieff P, Zhang L
    Cytokine, 2012-01-25;58(2):169-77.
    Species: Mouse
    Sample Types: Plasma
    Applications: Luminex Development
  6. Antigen-specific CD25- Foxp3- IFN-gamma(high) CD4+ T cells restrain the development of experimental allergic encephalomyelitis by suppressing Th17.
    Authors: Wildbaum G, Zohar Y, Karin N
    Am. J. Pathol., 2010-04-09;176(6):2764-75.
    Species: Mouse
    Sample Types: In Vivo, Whole Cells
    Applications: Neutralization
  7. Estrogen protects bone by inducing Fas ligand in osteoblasts to regulate osteoclast survival.
    Authors: Krum SA, Miranda-Carboni GA, Hauschka PV, Carroll JS, Lane TF, Freedman LP, Brown M
    EMBO J., 2008-01-24;27(3):535-45.
    Species: Mouse
    Sample Types: Whole Cells, Whole Tissue
    Applications: IHC-P, Neutralization

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Mouse Fas Ligand/TNFSF6 Antibody
By Anonymous on 01/31/2022
Application: WB Sample Tested: Osteoblasts Species: Mouse
Western Blot Mouse Fas Ligand/TNFSF6 Antibody MAB5262