Skip to main content Skip to navigation
Hamburger Menu
Search icon
Country Selector Country Selector
Middle East / Africa (English)
Products

Solutions

Discover & Learn

Resources & Tools

Support

Search icon Search icon
Close Menu
    Alert Icon
    Purified NA/LE Mouse Anti-Human CD178
    Purified NA/LE Mouse Anti-Human CD178
    Flow cytometric analysis of human CD178 on natural killer cells (CD16+). Peripheral blood mononuclear cells were cultured for 3 hours in media alone (right panel) or with ionomycin plus the metalloprotease inhibitor, KB8301 (left panel). KB8301 blocks CD178 cleavage resulting in high levels of cell surface CD178. The cells were stained with either Purified NA/LE Mouse Anti-Human CD178 (Cat. No. 556371; solid line histograms) or Purified Mouse IgG1, κ Isotype Control (Cat. No. 557273; dashed line histograms), followed by Biotin Goat Anti-Mouse Ig (Multiple Adsorption) (Cat. No. 550337). The cells were then incubated with normal mouse serum before adding FITC Mouse Anti-Human CD16 (Cat. No. 555406) and Streptavidin-PE (Cat. No. 554061). Fluorescent histograms were derived from gated events with the side and forward light-scattering characteristics of viable CD16+ cells. Flow cytometry was performed on a BD FACScan™ system.
    Purified NA/LE Mouse Anti-Human CD178
    Immunoprecipitation/western blot analysis of human CD178. CD178 was precipitated from human peripheral blood mononuclear cells (PBMC's) with either Purified NA/LE Mouse Anti-Human CD178 (Lane 1) or Purified Mouse IgG1, κ Isotype Control (Cat. No. 557273; Lane 2) and detected by western blot analysis with clone G247-4 (Cat. No. 556387). The bands above and below the 40 kDa CD178 band in lane 1 and lane 2 represent the heavy and light chain of IgG used for immunoprecipitation.
    Purified NA/LE Mouse Anti-Human CD178 Purified NA/LE Mouse Anti-Human CD178
    Flow cytometric analysis of human CD178 on natural killer cells (CD16+). Peripheral blood mononuclear cells were cultured for 3 hours in media alone (right panel) or with ionomycin plus the metalloprotease inhibitor, KB8301 (left panel). KB8301 blocks CD178 cleavage resulting in high levels of cell surface CD178. The cells were stained with either Purified NA/LE Mouse Anti-Human CD178 (Cat. No. 556371; solid line histograms) or Purified Mouse IgG1, κ Isotype Control (Cat. No. 557273; dashed line histograms), followed by Biotin Goat Anti-Mouse Ig (Multiple Adsorption) (Cat. No. 550337). The cells were then incubated with normal mouse serum before adding FITC Mouse Anti-Human CD16 (Cat. No. 555406) and Streptavidin-PE (Cat. No. 554061). Fluorescent histograms were derived from gated events with the side and forward light-scattering characteristics of viable CD16+ cells. Flow cytometry was performed on a BD FACScan™ system.
    Immunoprecipitation/western blot analysis of human CD178. CD178 was precipitated from human peripheral blood mononuclear cells (PBMC's) with either Purified NA/LE Mouse Anti-Human CD178 (Lane 1) or Purified Mouse IgG1, κ Isotype Control (Cat. No. 557273; Lane 2) and detected by western blot analysis with clone G247-4 (Cat. No. 556387). The bands above and below the 40 kDa CD178 band in lane 1 and lane 2 represent the heavy and light chain of IgG used for immunoprecipitation.
    Product Details
    Down Arrow Up Arrow


    BD Pharmingen™
    Fas ligand; FASL; FASLG; CD95 ligand; CD95L; CD95-L; TNFSF6; APTL; APT1LG1
    Human (QC Testing)
    Mouse IgG1
    Mouse T lymphoma cells (L5178Y) expressing human FasL
    Flow cytometry (Routinely Tested), Immunoprecipitation (Tested During Development), Neutralization (Reported), Western blot (Not Recommended)
    1.0 mg/ml
    356
    AB_396392
    No azide/low endotoxin: Aqueous buffered solution containing no preservative, 0.2µm sterile filtered. Endotoxin level is ≤0.01 EU/µg (≤0.001 ng/µg) of protein as determined by the LAL assay.
    RUO


    Preparation And Storage

    Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. This preparation contains no preservatives, thus it should be handled under aseptic conditions.
    Show More blue down arrow Show Less blue up arrow

    Recommended Assay Procedures

    NOK-1 has also been shown to neutralize the cytotoxic activity of FasL. Neutralization of FasL activity inhibits Fas-mediated killing. Purified NA/LE Mouse Anti-Human CD178 (Cat. No. 556371) should be used for all functional assays. NOK-1 and a related human FasL clone, NOK-2 [Cat. No. 556376 (purified) and No. 556375 (NA/LE)] may give different profiles in neutralization assays. It is thought that NOK-1 and NOK-2 likely recognize different FasL epitopes. Neither NOK-1 nor NOK-2 are suggested for western blot analysis.  

    Show More blue down arrow Show Less blue up arrow

    Product Notices

    1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
    2. An isotype control should be used at the same concentration as the antibody of interest.
    3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
    Show More blue down arrow Show Less blue up arrow
    556371 Rev. 4
    Antibody Details
    Down Arrow Up Arrow
    NOK-1

    The NOK-1 monoclonal antibody specifically recognizes CD178. Fas (CD95; APO-1) is a 45 kDa cell surface protein that mediates apoptosis when cross-linked with agonistic anti-Fas antibodies or by Fas ligand (FasL; CD178). Fas belongs to the TNF (Tumor Necrosis Factor)/NGF (Nerve Growth Factor) receptor family, and is expressed in various tissues and cells including the thymus, liver, ovary and lung. CD178 (FasL), a member of the TNF cytokine family, induces apoptosis by binding to Fas, its cell-surface receptor. FasL may exist as either membrane bound or soluble forms and is expressed by activated T and NK cells. FasL may also be constitutively expressed in some immunologically privileged sites, e.g., eye and testis. Fas and FasL play an important role in the induction of apoptosis, and thus regulate a variety of immunological responses.

    The NOK-1 antibody clone has been reported to recognize human FasL, recognizing both the membrane bound (FasL) and soluble (sFasL) forms. It is reported that the epitope for NOK-1 has been mapped to the COOH-terminus of FasL, at the region implicated in Fas binding. FasL and sFasL have been reported to migrate at reduced molecular weights of 40 and 26 kDa, respectively. However, the molecular weights observed in a particular sample may vary according to FasL and sFasL glycosylation and breakdown patterns as described in the literature.  The NOK-1 antibody clone is not recommended for the Western blot application.

    556371 Rev. 4
    Format Details
    Down Arrow Up Arrow
    NA/LE
    NA/LE refers to the culture and purification methods and buffer used to produce purified antibodies with no azide and low endotoxin: Aqueous buffered solution containing no preservative, 0.2µm sterile filtered. Endotoxin level is ≤0.01 EU/µg (≤0.001 ng/µg) of protein as determined by the LAL assay.NA/LE are perfectly suited to be used in culture or in vivo (for nonhuman studies) for functional assays — blocking, neutralizing, activation or depletion — where the presence of azide may damage cells or exogenous endotoxin may signal or activate cells.
    NA/LE
    556371 Rev.4
    Citations & References
    Down Arrow Up Arrow

    Development References (9)

    1. Kayagaki N, Kawasaki A, Ebata T, et al. Metalloproteinase-mediated release of human Fas ligand. J Exp Med. 1995; 182(6):1777-1783. (Immunogen: Flow cytometry, Neutralization). View Reference
    2. 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, Neutralization). View Reference
    3. Oyaizu N, Adachi Y, Hashimoto F, et al. Monocytes express Fas ligand upon CD4 cross-linking and induce CD4+ T cells apoptosis: a possible mechanism of bystander cell death in HIV infection. J Immunol. 1997; 158(5):2456-2463. (Clone-specific: Flow cytometry). View Reference
    4. Sieg S, Smith D, Yildirim Z, Kaplan D. Fas ligand deficiency in HIV disease. Proc Natl Acad Sci U S A. 1997; 94(11):5860-5865. (Clone-specific: Flow cytometry). View Reference
    5. 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
    6. 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
    7. Villunger A, Egle A, Marschitz I, et al. Constitutive expression of Fas (Apo-1/CD95) ligand on multiple myeloma cells: a potential mechanism of tumor-induced suppression of immune surveillance. Blood. 1997; 90(1):12-20. (Clone-specific: Flow cytometry, Neutralization). View Reference
    8. Walker PR, Saas P, Dietrich PY. Role of Fas ligand (CD95L) in immune escape: the tumor cell strikes back. J Immunol. 1997; 158(10):4521-4524. (Clone-specific: Neutralization). View Reference
    9. Zavazava N, Kronke M. Soluble HLA class I molecules induce apoptosis in alloreactive cytotoxic T lymphocytes. Nat Med. 1996; 2(9):1005-1010. (Clone-specific: Flow cytometry, Neutralization). View Reference
    View All (9) View Less
    556371 Rev. 4

    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.

    Website Feedback