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PE Mouse Anti-Human CD178
PE Mouse Anti-Human CD178
Flow cytometric analysis of CD178 (FasL) expression on FasL-transfected cells. Untransfected mouse L5187Y T lymphoma cells were cultured alone whereas human FasL-transfected L5187Y T lymphoma cells were cultured with the KB8301 metalloproteinase inhibitor for 24 hours. KB8301 blocks enzymatic cleavage of CD178 resulting in high CD178 levels to be expressed by the transfected cells. The untransfected (Left Panel) and transfected (Right Panel) cells were stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; dashed line histograms) or PE Mouse Anti-Human CD178 (Cat. No. 564261; solid line histograms). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Flow cytometric analysis of CD178 (FasL) expression on FasL-transfected cells. Untransfected mouse L5187Y T lymphoma cells were cultured alone whereas human FasL-transfected L5187Y T lymphoma cells were cultured with the KB8301 metalloproteinase inhibitor for 24 hours. KB8301 blocks enzymatic cleavage of CD178 resulting in high CD178 levels to be expressed by the transfected cells. The untransfected (Left Panel) and transfected (Right Panel) cells were stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; dashed line histograms) or PE Mouse Anti-Human CD178 (Cat. No. 564261; solid line histograms). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
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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)
5 µl
356
AB_2738713
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
564261 Rev. 2
Antibody Details
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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.

564261 Rev. 2
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
564261 Rev.2
Citations & References
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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
564261 Rev. 2

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