Anti-Fas Antibody

43 kDa

Fas/APO-1/CD95 (36 kDa) is a member of the tumor necrosis factor (TNF) receptor superfamily, a family of transmembrane receptors. Fas has been shown to be an important mediator of apoptotic cell death, as well as being involved in inflammation. Binding of the Fas ligand (Fas-L) induces trimerization of Fas in the target cell membrane. Activation of Fas causes the recruitment of Fas-associated protein with death domain (FADD) via interactions between the death domains of Fas and FADD. Procaspase 8 binds to Fas-bound FADD via interactions between the death effector domains (DED) of FADD and pro-caspase 8 leading to the activation of caspase 8. Activated caspase 8 cleaves (activates) other procaspases, in effect beginning a caspase cascade that ultimately leads to apoptosis. Caspases cleave nuclear lamins, causing the nucleus to break down and lose its normal structure. Fas-induced apoptosis can be effectively blocked at several stages by either FLICE-inhibitory protein (FLIP), by Bcl-2, or by the cytokine response modifier A (CrmA).

Biological Activity
The antibody demonstrates cytolytic activity on human cells that express Fas. Murine WR19L cells and L929 cells transfected with cDNA encoding human Fas undergo apoptosis in response to this antibody.

FS-7 (human diploid fibroblast cell line). Clone CH-11.

Detect Fas using this Anti-Fas Antibody (human, activating), clone CH11 has been published and validated for use in Flow Cytometry (FC), Immunocytochemistry (IC) and Western Blot (WB).

Research Category
Apoptosis & Cancer

Research Sub Category
Apoptosis - Additional

Western Blot:
0.5-2 μg/mL of a previous lot detected Fas in a Raji cell lysate.
Immunocytochemistry:
5-10 μg/mL of a previous lot detected Fas on HeLa cells fixed with 4% formalin/2% acetic acid.
Flow cytometry:
A previous lot of was tested by an independent laboratory using 20 μg/mL of anti-Fas, clone CH11 (Yonehara, S., 1989; Kobayashi, N., 1990).

This antibody does not recognize TNF, and does not cross-react with mouse Fas. Fas ligand will induce apoptosis in human, mouse and rat systems.

This antibody recognizes the human cell surface antigen Fas, Mr 43 kDa expressed in various human cells, including myeloid cells, T lympho-blastoid cells, and diploid fibroblasts.

Immunoaffinity Chromatography

Purified mouse monoclonal IgM in buffer containing PBS, pH 7.2, with 50% glycerol. Liquid at -20ºC.

Stable for 1 year at -20°C from date of receipt. For maximum recovery of the product, centrifuge the original vial prior to removing the cap.

Control
Human liver tumor, human breast tumor or Jurkat whole cell lysate, Raji cell lysate.

Routinely evaluated by demonstrating cytolytic activity on human cells that express Fas. Murine WR19L cells and L929 cells transfected with cDNA encoding human Fas undergo apoptosis in response to this antibody.

Apoptosis Assay Analysis:
15-20 µg/mL of this lot maximally induced apoptosis of human Jurkat cells with 83% mortality after 24 hours of treatment.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

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