FAS anticorps

N° du produit ABIN1177357

Cet anticorps Souris Monoclonal détecte spécifiquement FAS dans FACS et Func. Il présente une réactivité envers Humain, Porc, Macaque rhésus, Chien, Babouin et Cynomolgus et a été mentionné dans 2+ publications.

Aperçu rapide pour FAS anticorps (ABIN1177357)

Antigène: FAS (TNF Receptor Superfamily, Member 6 (FAS))

Reactivité: Humain, Porc, Macaque rhésus, Chien, Babouin, Cynomolgus

Hôte: Souris

Clonalité: Monoclonal

Conjugué: Cet anticorp FAS est non-conjugé

Application: Flow Cytometry (FACS), Functional Studies (Func)

Clone: DX2

Détail du produit anti-FAS anticorps

Marque: BD Pharmingen™

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Stérilité: 0.2 μm filtered

niveau d'endotoxine: Endotoxin level is ≤ 0.01 EU/μg (≤ 0.001 ng/μg) of protein as determined by the LAL assay.

Immunogène: Human CD95-transfected L ceils

Isotype: IgG1 kappa

Information d'application

Protocole: 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

Stockage

Format: Liquid

Concentration: 1.0 mg/mL

Buffer: No azide/low endotoxin: Aqueous buffered solution containing no preservative, 0.2μm sterile filtered.

Agent conservateur: Azide free

Stock: 4 °C

Stockage commentaire: Store undiluted at 4°C. This preparation contains no preservatives, thus it should be handled under aseptic conditions.

Références pour anticorps anti-FAS (ABIN1177357)

Cifone, De Maria, Roncaioli, Rippo, Azuma, Lanier, Santoni, Testi: "Apoptotic signaling through CD95 (Fas/Apo-1) activates an acidic sphingomyelinase." dans: The Journal of experimental medicine, Vol. 180, Issue 4, pp. 1547-52, (1994) (PubMed).

Itoh, Yonehara, Ishii, Yonehara, Mizushima, Sameshima, Hase, Seto, Nagata: "The polypeptide encoded by the cDNA for human cell surface antigen Fas can mediate apoptosis." dans: Cell, Vol. 66, Issue 2, pp. 233-43, (1991) (PubMed).

Détails sur FAS

Antigène: FAS (TNF Receptor Superfamily, Member 6 (FAS))

Autre désignation: CD95

Sujet: 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.
Synonyms: Fas/APO-1

Pathways: Signalisation p53, Apoptose, Production of Molecular Mediator of Immune Response, Positive Regulation of Endopeptidase Activity