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Purified Mouse Anti-Human CD178
Purified Mouse Anti-Human CD178

Western blot analysis of Fas Ligand (FasL). L5187Y human T lymphoma cells were transfected with human FasL cDNA and then not treated (lane 1) or treated (lane 2) with the metalloproteinase inhibitor, KB8301. KB8301 blocks FasL cleavage from the cell surface, resulting in high levels membrane expressed FasL. The blot shows that whereas FasL is detected in untreated cells (lane 1), levels increased dramatically when cells were treated with KB8301 (lane 2). A mouse IgG1 isotype matched antibody was used as a negative control (lane 3).

Western blot analysis of Fas Ligand (FasL). L5187Y human T lymphoma cells were transfected with human FasL cDNA and then not treated (lane 1) or treated (lane 2) with the metalloproteinase inhibitor, KB8301. KB8301 blocks FasL cleavage from the cell surface, resulting in high levels membrane expressed FasL. The blot shows that whereas FasL is detected in untreated cells (lane 1), levels increased dramatically when cells were treated with KB8301 (lane 2). A mouse IgG1 isotype matched antibody was used as a negative control (lane 3).

Product Details
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BD Pharmingen™
Fas Ligand, CD95 Ligand
Human (QC Testing)
Mouse IgG1
Recombinant Human FasL
Western blot (Routinely Tested), Immunohistochemistry-frozen, Immunoprecipitation (Tested During Development), Flow cytometry (Not Recommended)
42 kDa, 40 kDa (membrane); 26 kDa (soluble)
0.5 mg/ml
AB_396402
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.

Recommended Assay Procedures

Applications include immunoprecipitation (1-2 µg/ml), western blot analysis (1-2 µg/ml) and immunohistochemical staining of acetone-fixed frozen tissue sections (0.5-4 µg/ml). G247-4 is not recommended for flow cytometry. For flow cytometry application, clone NOK-1 (purified, Cat. No. 556372; or biotin-conjugated, Cat. No. 556374) is recommended.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
556387 Rev. 10
Antibody Details
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G247-4

Fas (APO-1, CD95) is a 45 kD cell surface protein that mediates apoptosis when crosslinked with agonistic anti-Fas antibodies or Fas ligand (FasL). Fas belongs to the TNF (tumor necrosis factor)/NGF (nerve growth factor) receptor family, and is expressed in various tissue and cells including the thymus, liver, ovary and lung. FasL is a member of the TNF cytokine family that induces apoptosis by binding to Fas, its cell-surface receptor. FasL may exist in both membrane and soluble forms and expressed on activated T cells, NK cells, and other "immunologically privileged" sites. Both Fas and FasL are thought to play an important role in the apoptotic processes that take place during T cell development.

G247-4 recognizes human FasL. It recognizes both the membrane bound (FasL) and soluble (sFasL) forms. A recombinant protein containing the external domain of human FasL was used as immunogen. FasL and sFasL migrate at reduced  molecular weights of 40 and 26 kD, respectively. However, the molecular weights observed in a particular sample may vary according to FasL and sFasL glycosylation and breakdown patterns as described in Tanaka et al.  For example, FasL may migrate as a doublet of 40 and 42 kD.

556387 Rev. 10
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
556387 Rev.10
Citations & References
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Development References (7)

  1. Griffith TS, Ferguson TA. The role of FasL-induced apoptosis in immune privilege. Immunol Today. 1997; 18(5):240-244. (Biology). View Reference
  2. Kayagaki N, Kawasaki A, Ebata T, et al. Metalloproteinase-mediated release of human Fas ligand. J Exp Med. 1995; 182(6):1777-1783. (Biology). View Reference
  3. Orlinick JR, Elkon KB, Chao MV. Separate domains of the human fas ligand dictate self-association and receptor binding. J Biol Chem. 1997; 272(51):32221-32229. (Clone-specific: Immunoprecipitation, Western blot). View Reference
  4. Orlinick JR, Vaishnaw A, Elkon KB, and Chao MV. Requirement of cysteine-rich repeats of the Fas receptor for binding by the Fas ligand. J Biol Chem. 1997; 272:28889-28894. (Biology).
  5. Sträter J, Wellisch I, Riedl S. CD95 (APO-1/Fas)-mediated apoptosis in colon epithelial cells: a possible role in ulcerative colitis. Gastroenterology. 1997; 113(1):160-167. (Clone-specific: Immunoprecipitation, Western blot). View Reference
  6. Takahashi T, Tanaka M, Brannan CI, et al. Generalized lymphoproliferative disease in mice, caused by a point mutation in the Fas ligand. Cell. 1994; 76(6):969-976. (Biology). View Reference
  7. Tanaka M, Suda T, Takahashi T, Nagata S. Expression of the functional soluble form of human Fas ligand in activated lymphocytes. EMBO J. 1995; 14(6):1129-1135. (Biology). View Reference
View All (7) View Less
556387 Rev. 10

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.