SPECIFICITY
The Kay-10 antibody reacts with Fas Ligand (FasL, CD95L) expressed on activated T lymphocytes of selected strains of mice (e.g., C57BL/6, C3H, MRL, NOD, NZB, and SJL, but not BALB/c, DBA/1, nor DBA/2).1 It also reacts with Cos cells transfected with mFasL cDNA derived from C57BL/6 and C3H mice, but not with mFasL cDNA from BALB/c nor DBA/2 mice.1 In the mouse, FasL is expressed on activated T cell lines2,3,4,5 and in spleen, testis, and eye.6 FasL mRNA has been demonstrated at various levels in bone marrow, thymus, spleen, lymph node, lung, small intestine, testis, and uterus.6,7,8 Moreover, T-cell activators, but not B-cell activators, enhanced the expression of FasL mRNA in splenocytes; and FasL mRNA was restricted to the T-cell lineage among a panel of cell lines from lymphoid tissues.8 FasL is not functional in mice homozygous for the gld (generalized lymphoproliferative disease) mutation; these mice cannot limit the expansion of activated lymphocytes and develop autoimmune disease.9 FasL is a type II membrane protein, a member of the TNF/NGF family, which binds to CD95 (Fas).10 FasL and its counter-receptor CD95 are believed to participate in T-cell development, the regulation of immune responses, and cell-mediated cytotoxic mechanisms.3-13 It has been demonstrated that human FasL is released from the surface of transfectant cells and activated human T cells by metalloproteinases.14
USAGE
The antibody has been tested by immunofluorescent staining, in the presence of metalloproteinase inhibitor, of mFasL-transfected and activated T cell lines (1 - 4 µg/million cells) with flow cytometric analysis (for information on how to obtain the metalloproteinase inhibitor, please [E-Mail] us. Because FasL is expressed at low density on activated cells, we recommend the use of a biotinylated second-step antibody with a "bright" third step reagent, such as Streptavidin-PE (Cat. no. 13025D). Please note that the conditions and kinetics of cell-surface expression of the FasL protein on activated normal T cells (not cell lines) have not yet been published and are currently under evaluation.1 Other reported applications include blocking of the cytotoxic activity of the mFasL-transfected L5178Y T lymphoma.1 For more details, please contact our Technical Services Department. Since applications vary, each investigator must determine dilutions appropriate for individual use.
REFERENCES
1. Kayagaki, N. Personal communication.
2. Ramsdell, F., M.S. Seaman, R.E. Miller, K.S. Picha, M.K. Kennedy, and D.H. Lynch. 1994. Differential ability of Th1 and Th2 T cells to express Fas ligand and to undergo activation-induced cell death.
Int. Immunol.
6: 1545 - 1553.
3. Brunner, T., R.J. Mogil, D. LaFace, N.J. Yoo, A. Mahboubi, F. Echeverri, S. J. Martin, W.R. Force, D.H. Lynch, C.F. Ware, and D.R. Green. 1995. Cell-autonomous Fas (CD95)/Fas-ligand interaction mediates activation-induced apoptosis in T-cell hybridomas.
Nature
373: 441 - 444.
4. Ju, S.-T., D.J. Panka, H. Cui, R. Ettinger, M. El-Khatib, D.H. Sherr, B.Z. Stanger, A. Marshak-Rothstein. 1995. Fas(CD95)/FasL interactions required for programmed cell death after T-cell activation.
Nature
373: 444 - 448.
5. Vignaux, F., E. Vivier, B. Malissen, V. Depraetere, S. Nagata, and P. Golstein. 1995. TCR/CD3 coupling to Fas-based cytotoxicity.
J. Exp. Med.
181: 781 - 786.
6. Griffith, T.S., T. Brunner, S.M. Fletcher, D.R. Green, and T.A. Ferguson. 1995. Fas ligand-induced apoptosis as a mechanism of immune privilege.
Science
270: 1189 - 1192.
7. Bellgrau, D., D. Gold, H. Selawry, J. Moore, A. Franzusoff, and R.C. Duke. 1995. A role for CD95 ligand in preventing graft rejection.
Nature
377: 630 - 632.
8. Suda, T., T. Okazaki, Y. Naito, T. Yokota, N. Arai, S. Ozaki, K. Nakao, and S. Nagata. Expression of the Fas ligand in cells of T cell lineage. 1995.
J. Immunol.
154: 3806 - 3813.
9. Takahashi, T., M. Tanaka, C.I. Brannan, N.A. Jenkins, N.G. Copeland, T. Suda, and S. Nagata. 1994. Generalized lymphoproliferative disease in mice, caused by a point mutation in the Fas ligand.
Cell
76: 969 - 976.
10. Smith, C.A., T. Farrah, and R.G. Goodwin. 1994. The TNF receptor superfamily of cellular and viral proteins: Activation, costimulation, and death.
Cell
76: 959 - 962.
11. Kojima, H., N. Shinohara, S. Hanaoka, Y. Someya-Shirota, Y. Takagaki, H. Ohno, T. Saito, T. Katayama, H. Yagita, K. Okumura, Y. Shinkai, F.W. Alt, A. Matsuzawa, S. Yonehara, and H. Takayama. 1994. Two distinct pathways of specific killing revealed by perforin mutant cytotoxic T lymphocytes.
Immunity
1: 357 - 364.
12. Lynch, D.H., F. Ramsdell, and M.R. Alderson. 1995. Fas and FasL in the homeostatic regulation of immune responses.
Immunol. Today
16: 569 - 574.
13. Lau, H.T., M. Yu, A. Fontana, C.J. Stoeckert, Jr. 1996. Prevention of islet allograft rejection with engineered myoblasts expressing FasL in mice.
Science
273: 109 - 112.
14. Kayagaki, N., A. Kawasaki, T. Ebata, H. Ohmoto, S. Ikeda, S. Inoue, K. Yoshino, K. Okumura, and H. Yagita. 1995. Metalloproteinase-mediated release of human Fas Ligand.
J. Exp. Med.
182: 1777 - 1783.
| Cat. No. | Description | Clone | Isotype | Size |
| 09931D | Purified anti-mouse Fas Ligand | KAY-10 | Mouse IgG2b | 0.5mg |
Related products for apoptosis research include antibodies to CD95 (FAS), 4-1BB, CD30. We also offer the APO-DIRECT™ Kit for single-step detection of apoptotic cells by flow cytometry.
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