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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H | FC, ICC, WB | M | Affinity Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | 05-201 |
| Brand Family | Upstate |
| Trade Name |
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| Description | Anti-Fas Antibody (human, activating), clone CH11 |
| Alternate Names |
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| Background Information | 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. |
| Product Information | |
|---|---|
| Format | Affinity Purified |
| Control |
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| Presentation | Purified mouse monoclonal IgM in buffer containing PBS, pH 7.2, with 50% glycerol. Liquid at -20ºC. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| 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). |
| Key Applications |
|
| Application Notes | 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). |
| Biological Information | |
|---|---|
| Immunogen | FS-7 (human diploid fibroblast cell line). Clone CH-11. |
| Clone | CH11 |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Mouse |
| 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. |
| Isotype | IgM |
| Species Reactivity |
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| Species Reactivity Note | Proven to react with human. 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. |
| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | The protein encoded by this gene is a member of the TNF-receptor superfamily. This receptor contains a death domain. It has been shown to play a central role in the physiological regulation of programmed cell death, and has been implicated in the pathogenesis of various malignancies and diseases of the immune system. The interaction of this receptor with its ligand allows the formation of a death-inducing signaling complex that includes Fas-associated death domain protein (FADD), caspase 8, and caspase 10. The autoproteolytic processing of the caspases in the complex triggers a downstream caspase cascade, and leads to apoptosis. This receptor has been also shown to activate NF-kappaB, MAPK3/ERK1, and MAPK8/JNK, and is found to be involved in transducing the proliferating signals in normal diploid fibroblast and T cells. At least eight alternatively spliced transcript variants encoding seven distinct isoforms have been described. The isoforms lacking the transmembrane domain may negatively regulate the apoptosis mediated by the full length isoform. |
| Gene Symbol |
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| Purification Method | Immunoaffinity Chromatography |
| UniProt Number |
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| UniProt Summary | FUNCTION: SwissProt: P25445 # Receptor for TNFSF6/FASLG. The adapter molecule FADD recruits caspase-8 to the activated receptor. The resulting death- inducing signaling complex (DISC) performs caspase-8 proteolytic activation which initiates the subsequent cascade of caspases (aspartate-specific cysteine proteases) mediating apoptosis. FAS- mediated apoptosis may have a role in the induction of peripheral tolerance, in the antigen-stimulated suicide of mature T-cells, or both. The secreted isoforms 2 to 6 block apoptosis (in vitro). SIZE: 335 amino acids; 37732 Da SUBUNIT: Binds DAXX. Interacts with HIPK3. Part of a complex containing HIPK3 and FADD (By similarity). Binds RIPK1 and FAIM2. Interacts with BRE. SUBCELLULAR LOCATION: Isoform 1: Cell membrane; Single-pass type I membrane protein. & Isoform 2: Secreted. & Isoform 3: Secreted. & Isoform 4: Secreted. & Isoform 5: Secreted. & Isoform 6: Secreted. TISSUE SPECIFICITY: Isoform 1 and isoform 6 are expressed at equal levels in resting peripheral blood mononuclear cells. After activation there is an increase in isoform 1 and decrease in the levels of isoform 6. DOMAIN: SwissProt: P25445 Contains a death domain involved in the binding of FADD, and maybe to other cytosolic adapter proteins. DISEASE: SwissProt: P25445 # Defects in FAS are the cause of autoimmune lymphoproliferative syndrome type 1A (ALPS1A) [MIM:601859]; also known as Canale-Smith syndrome (CSS). ALPS is a childhood syndrome involving hemolytic anemia and thrombocytopenia with massive lymphadenopathy and splenomegaly. SIMILARITY: Contains 1 death domain. & Contains 3 TNFR-Cys repeats. |
| Molecular Weight | 43 kDa |
| Product Usage Statements | |
|---|---|
| Quality Assurance | 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. |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | 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. |
| Packaging Information | |
|---|---|
| Material Size | 50 µg |