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05-201

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

clone CH11, Upstate®, from mouse

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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

biological source

mouse

Quality Level

antibody form

affinity purified immunoglobulin

antibody product type

primary antibodies

clone

CH11, monoclonal

purified by

affinity chromatography

species reactivity

human

manufacturer/tradename

Upstate®

technique(s)

flow cytometry: suitable
immunocytochemistry: suitable
western blot: suitable

isotype

IgM

NCBI accession no.

UniProt accession no.

shipped in

dry ice

target post-translational modification

unmodified

Gene Information

human ... FAS(355)

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This Item
05-338F4424SAB5700786
antibody form

affinity purified immunoglobulin

antibody form

purified immunoglobulin

antibody form

purified from hybridoma cell culture

antibody form

affinity isolated antibody

clone

CH11, monoclonal

clone

ZB4, monoclonal

clone

DX2, monoclonal

clone

polyclonal

species reactivity

human

species reactivity

human

species reactivity

human

species reactivity

rat, human, mouse

manufacturer/tradename

Upstate®

manufacturer/tradename

Upstate®

manufacturer/tradename

-

manufacturer/tradename

-

technique(s)

flow cytometry: suitable, immunocytochemistry: suitable, western blot: suitable

technique(s)

flow cytometry: suitable, neutralization: suitable, western blot: suitable

technique(s)

flow cytometry: 4-20 μg/mL using cultured human Burkitt’s lymphoma Raji cells, microarray: suitable

technique(s)

western blot: 1:500-1:1000

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 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.

Immunogen

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

Application

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).

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

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

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


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J B Mannick et al.
Science (New York, N.Y.), 284(5414), 651-654 (1999-04-24)
Only a few intracellular S-nitrosylated proteins have been identified, and it is unknown if protein S-nitrosylation/denitrosylation is a component of signal transduction cascades. Caspase-3 zymogens were found to be S-nitrosylated on their catalytic-site cysteine in unstimulated human cell lines and
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
Considering Fas ligand as a target for therapy.
Linkermann, Andreas, et al.
Expert Opinion on Therapeutic Targets, 9, 119-134 (2005)
F Basolo et al.
Laboratory investigation; a journal of technical methods and pathology, 80(9), 1413-1419 (2000-09-27)
The Fas-FasL system seems to mediate thyrocyte death in Hashimoto's thyroiditis. In thyroid cancer, down-regulation of bcl-2 seems to alter apoptosis control. We compared the expression of immunoreactive Fas and FasL in normal thyroid with that of tumors ranging from

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Application note on how the CellASIC® ONIX2 microfluidic system can be used to analyze caspase-3 mediated apoptosis/cell death and cellular hypoxia in live immune and cancer cell lines.

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