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05-201

Sigma-Aldrich

Anti-Fas Antibody (human, activating), clone CH11

clone CH11, Upstate®, from mouse

Synonym(s):
APO-1 cell surface antigen, CD95 antigen, Fas (TNF receptor superfamily, member 6), Fas AMA, Fas antigen, apoptosis antigen 1, tumor necrosis factor receptor superfamily, member 6
eCl@ss:
32160702
NACRES:
NA.41

Quality Level

biological source

mouse

antibody form

affinity purified immunoglobulin

antibody product type

primary antibodies

clone

CH11, monoclonal

purified by

affinity chromatography

species reactivity

human

manufacturer/tradename

Upstate®

application(s)

flow cytometry: suitable
immunocytochemistry: suitable
western blot: suitable

isotype

IgM

NCBI accession no.

UniProt accession no.

shipped in

dry ice

General description

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.

Specificity

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.
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.

Immunogen

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

Application

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).
Research Sub Category
Apoptosis - Additional
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

Quality

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.

Target description

43 kDa

Physical form

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

Storage and Stability

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.

Analysis Note

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

Other Notes

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

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 2

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis

Certificate of Origin

S DeFranco et al.
Diabetes, 50(3), 483-488 (2001-03-15)
Fas (CD95) triggers programmed cell death and is involved in cell-mediated cytotoxicity and in shutting off the immune response. Inherited loss-of-function mutations hitting the Fas system cause the autoimmune/lymphoproliferative syndrome (ALPS). We have recently shown that ALPS patients' families display...
K Lamberth et al.
Tissue antigens, 58(3), 171-180 (2001-11-13)
Early apoptosis in Jurkat T-lymphoma cells was induced by agonistic anti-Fas Ab or by anisomycin which activates the stress kinases SAPK/JNK. Apoptosis was inhibited by ligation of major histocompatibility complex class I antigens (MHC-I). MHC-I ligation induced upregulation of the...
K Wright et al.
The Journal of biological chemistry, 274(24), 17193-17201 (1999-06-08)
A combination of the pro-inflammatory cytokines interleukin (IL)-1alpha, interferon (IFN)-gamma, and tumor necrosis factor (TNF)-alpha induces nitric oxide synthase mRNA expression and nitric oxide (NO) generation in the human colon carcinoma cell line HT-29. This can be inhibited by pretreatment...
A L Kim et al.
The Journal of biological chemistry, 274(49), 34924-34931 (1999-11-27)
A p53-derived C-terminal peptide induced rapid apoptosis in breast cancer cell lines carrying endogenous p53 mutations or overexpressed wild-type (wt) p53 but was not toxic to nonmalignant human cell lines containing wt p53. Apoptosis occurred through a Fas/APO-1 signaling pathway...
M MacFarlane et al.
The Journal of cell biology, 148(6), 1239-1254 (2000-03-22)
Tumor necrosis factor-related apoptosis- inducing ligand (TRAIL) -induced apoptosis, in transformed human breast epithelial MCF-7 cells, resulted in a time-dependent activation of the initiator caspases-8 and -9 and the effector caspase-7. Cleavage of caspase-8 and its preferred substrate, Bid, preceded...

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