|Antigen||Fas (TNF Receptor Superfamily, Member 6) (FAS) Antibodies|
|Reactivity||Baboon, Cynomolgus, Dog (Canine), Human, Pig (Porcine), Rhesus Monkey Alternatives|
|Conjugate||This FAS antibody is un-conjugated Alternatives|
Functional Studies (Func), Flow Cytometry (FACS)
|2 references available|
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Product Details anti-FAS AntibodyTarget Details FAS Application Details Handling References for anti-FAS antibody (ABIN1177357) Images
|Purification||The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.|
|Sterility||0.2 μm filtered|
|Endotoxin Level||Endotoxin level is ≤ 0.01 EU/μg (≤ 0.001 ng/μg) of protein as determined by the LAL assay.|
|Immunogen||Human CD95-transfected L ceils|
Target Details FASProduct Details anti-FAS Antibody Application Details Handling References for anti-FAS antibody (ABIN1177357) Images back to top
|Alternative Name||CD95 (FAS Antibody Abstract)|
Reacts with the 45 kDa transmembrane cell surface Fas or Apo-1 antigen (designated CD95 at the Fifth HLDA Workshop) which is expressed on a variety of normal and neoplastic cells. The Fas/Apo-1 antigen is a polypeptide, which plays a role in the programmed sequence of events leading to cell death, termed apoptosis. The DX2 clone specifically reacts with murine L cells, murine L1210 leukemia cells and murine P815 mastocytoma cells transfected with human Fas cDNA but not with untransfected parental cell lines. Cross-linking with DX2 delivers an apoptotic signal indicating that DX2 recognizes a functional epitope of the CD95 antigen.
|Research Area||Surface Receptors of Immune Cells|
|Pathways||p53 Signaling, Apoptosis, Production of Molecular Mediator of Immune Response, Positive Regulation of Endopeptidase Activity|
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1. Maintain cells in culture with 10% FBS medium, change the medium one day before starting the induction of apoptosis. Harvest and pellet the cells, resuspend in 10% FBS medium at a density of 0.5-1.0 x 10^6 cells/ml.
2. In a 96-well microtiter plate, add 2.5 µg/50 µl CD95 NA/LE™, 0.5 µg/50 µl rProt G, 0.5-1.0 x 10^5 cells and 10% FBS medium to a total volume of 200 µl. Negative controls should consist of: 1) 0.5-1.0 x 10^5 cells with 10% FBS medium alone (200 µl total volume), and 2) 0.5-1.0 x 10^5 cells with 0.5 µg/50 µl rProtG and 10% FBS medium.
3. Incubate the 96-well plate at 37°C, 5% CO2, for 12-24 hours. Traditionally, apoptosis has been observed by light microscopy, MTT, or gel electrophoresis (DNA fragmentation). Investigators may be interested in several products offered by BD Biosciences that may be used for the detection of apoptotic cells by flow cytometry: ApoDirect™ Kit (Cat. No. 556381), ApoBRDU™ Kit (Cat. No. 556405) and Annexin V-FITC (Cat. No. 556420, Cat. No. 556419). Because these methods vary in sensitivity, it may be necessary to titer the rProtG and/or CD95 NA/LE™ to obtain optimal results.
Notes: The suggestions given in this procedure are based on conditions that were found to be optimal for the induction of apoptosis by anti-human CD95 (Fas). Studies have shown that the addition of protein G can significantly enhance the efficiency of DX2 in this type of functional assay. It is important to note that there is a great deal of variation between cell lines in the level of apoptosis that can be induced through the Fas receptor. It has been reported that, although some cells express the Fas antigen, they do not necessarily undergo Fas-mediated apoptosis (i.e., the Fas antigen expressed may be anti-Fas sensitive or insensitive). Daudi, Jurkat, and HPB-ALL cells are good positive controls as they are strongly induced by anti-Fas to undergo apoptosis.
|Restrictions||For Research Use only|
HandlingProduct Details anti-FAS Antibody Target Details FAS Application Details References for anti-FAS antibody (ABIN1177357) Images back to top
|Buffer||No azide/low endotoxin: Aqueous buffered solution containing no preservative, 0.2μm sterile filtered.|
|Storage Comment||Store undiluted at 4°C. This preparation contains no preservatives, thus it should be handled under aseptic conditions.|
References for anti-FAS antibody (ABIN1177357)Product Details anti-FAS Antibody Target Details FAS Application Details Handling Images back to top
|Product cited in:||
Cifone, De Maria, Roncaioli, Rippo, Azuma, Lanier, Santoni, Testi: "Apoptotic signaling through CD95 (Fas/Apo-1) activates an acidic sphingomyelinase." in: The Journal of experimental medicine, Vol. 180, Issue 4, pp. 1547-52, 1994
Itoh, Yonehara, Ishii, Yonehara, Mizushima, Sameshima, Hase, Seto, Nagata: "The polypeptide encoded by the cDNA for human cell surface antigen Fas can mediate apoptosis." in: Cell, Vol. 66, Issue 2, pp. 233-43, 1991
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