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

Immunoprecipitation of Fas Ligand (FasL). T lymphoma L5187Y cells were transfected with human FasL cDNA and treated or not treated with metalloprotease inhibitor KB8301. KB8301 blocks FasL cleavage resulting in high levels of cell surface FasL. FasL is detected in S-labeled cells treated with inhibitor KB8301 (lane 1), but not in untreated cells (lane 3). Isotype (negative) controls are shown in lanes 2 and 4. Purified NOK-2 (Cat. No. 556376) or mouse IgG2a (Cat. No. 553454) was used for immunoprecipitations.  

Immunoprecipitation of Fas Ligand (FasL). T lymphoma L5187Y cells were transfected with human FasL cDNA and treated or not treated with metalloprotease inhibitor KB8301. KB8301 blocks FasL cleavage resulting in high levels of cell surface FasL. FasL is detected in S-labeled cells treated with inhibitor KB8301 (lane 1), but not in untreated cells (lane 3). Isotype (negative) controls are shown in lanes 2 and 4. Purified NOK-2 (Cat. No. 556376) or mouse IgG2a (Cat. No. 553454) was used for immunoprecipitations.  

Product Details
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BD Pharmingen™
Fas Ligand, CD95 Ligand
Human (QC Testing)
Mouse IgG2a
L51788Y Mouse T Lymphoma Cells Expressing Recombinant Human FasL
Immunoprecipitation (Routinely Tested), Flow cytometry (Tested During Development), ELISA, Neutralization (Reported), Western blot (Not Recommended)
40, 26 kDa
0.5 mg/ml
AB_396397
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

IP: Applications include immunoprecipitation (1-2 µg/one million cells).

Functional Assay: NOK-2 has also been shown to neutralize the cytotoxic activity of FasL. Neutralization of FasL activity inhibits Fas-mediating  killing. The NA/LE format of NOK-2 (Cat. No. 556375) should be used for all functional assays. NOK-2 and a related human FasL clone, NOK-1 [Cat. No. 556371 (NA/LE)] may give different profiles in neutralization assays. It is thought that NOK-1 and NOK-2 likely recognize different FasL epitopes.

Flow Cytometry: Another clone,  NOK-1, (556372, 556373)  are  recommended for flow cytometry. NOK-2 appears weaker for flow cytometry than NOK-1.

Western Blot: Neither NOK-1 nor NOK-2 are suggested for western blot analysis. Another human FasL clone, G247-4 (Cat. No. 556387) is suggested for the western blot analysis.

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.
556376 Rev. 6
Antibody Details
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NOK-2

Fas (APO-1, CD95) is a 45 kDa cell surface protein that mediates apoptosis when cross-linked 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 40kDa TNF family membrane protein that induces apoptosis by binding to Fas, its cell-surface receptor. FasL is expressed on activated T and NK cells. Both Fas and FasL are thought to play an important role in  the apoptotic processes that take place during T cell development.

NOK-2 recognizes human FasL. It recognizes both the membrane bound (FasL) and soluble (sFasL) forms. L5178Y mouse T lymphoma cells expressing recombinant human FasL were used as immunogen. FasL and sFasL 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 Tanaka et al.

556376 Rev. 6
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
556376 Rev.6
Citations & References
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Development References (3)

  1. Kayagaki N, Kawasaki A, Ebata T, et al. Metalloproteinase-mediated release of human Fas ligand. J Exp Med. 1995; 182(6):1777-1783. (Clone-specific: ELISA, Immunoprecipitation, Neutralization). View Reference
  2. 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
  3. 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
556376 Rev. 6

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.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.