FAS recombinant monoclonal antibody, clone R-125224
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More Files
- More Functions
Western Blot
Western Blot analysis were resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit IgG version of R-125224 at 2 ug/mL before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence. The expected running size for unmodified Fas is 37.7kDa, but this protein is glycosylated at several positions leading to the observed running size.
Immunofluorescence
Immunofluorescence analysis of paraformaldehyde fixed MCF7 cells permeabilized with 0.15% Triton and stained with the chimeric mouse IgG1 version of R-125224 at 10 ug/mL for 1h followed by Alexa Fluor 488 secondary antibody (2 ug/mL), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom, DAPI, merged channels and an isotype control. The isotype control was stained with an anti-unknown specificity antibody followed by Alexa Fluor 488 secondary antibody.
Flow Cytometry
Flow-cytometry were stained with unimmunized rabbit IgG antibody (black line) or the rabbit IgG-chimeric version of R-125224 (blue line) at a dilution of 1:100 for 1h at RT. After washing, bound antibody was detected using a goat anti-rabbit IgG AlexaFluor 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.
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Specification
Product Description
Rabbit recombinant monoclonal antibody raised against human R-125224.
Antibody Species
Rabbit
Immunogen
Original antibody is raised against by the humanization of the murine HFE7A anti-Fas antibody by grafting the CDR regions to the framework regions of the human 8E10 antibody and substituting key framework residues from the murine antibody into the 8E10 sequence. The original HFE7A was derived from a hybridoma cell line generated by the fusion of NS1 myeloma cells with splenocytes from Fasdeficient mice which had been immunized with partially purified recombinant human Fas-AIC2A chimera protein consisting of the extracellular region of human Fas antigen (aa -16 to 150) and the extracellular region of the murine IL-3 receptor AIC2 (aa 3-423). The HFE7A hybridoma was selected after screening by flow cytometry for the production of antibodies with the ability to bind to the WR19L12a transformed murine T cell lymphoma cell line expressing human Fas or the L5178YA1 cell line expressing murine Fas, but not to the parental WR19L or L5178Y cell.
Reactivity
Human
Specificity
R-125224 binds to the extracellular portion of human Fas at an eptiope consisting of the sequence RTQNTKCRCK (aa 105-114) (pmid: 11754745). Fas is a type I membrane protein which belongs to the tumor necrosis factor (TNF) receptor/nerve growth factor (NGF) receptor superfamily. It is able to transduce apoptotic signals into the cell when bound by its ligand FasL (Fas ligand), which is primarily expressed in activated T lymphoid-myeloid lineage cells, in the eye, in reproductive organs and in some tumors. The Fas-FasL system is known to play an important role in maintaining the immune system as mice with Fas-defective lymphoproliferation (lpr) and FasL-defective generalized lymphoproliferative disease (gld) mutations develop massive lymphadenopathy and autoimmune diseases.
Form
Liquid
Purification
Protein A affinity purification
Isotype
IgG, Kappa
Recommend Usage
Flow cytometry
Immunofluorescence
Western Blot
The optimal working dilution should be determined by the end user.Storage Buffer
In PBS with 0.02% Proclin 300
Storage Instruction
Store at 4°C for up to 3 months. For longer storage, aliquot and store at -20°C.
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Applications
Western Blot
Western Blot analysis were resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit IgG version of R-125224 at 2 ug/mL before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence. The expected running size for unmodified Fas is 37.7kDa, but this protein is glycosylated at several positions leading to the observed running size.Immunofluorescence
Immunofluorescence analysis of paraformaldehyde fixed MCF7 cells permeabilized with 0.15% Triton and stained with the chimeric mouse IgG1 version of R-125224 at 10 ug/mL for 1h followed by Alexa Fluor 488 secondary antibody (2 ug/mL), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom, DAPI, merged channels and an isotype control. The isotype control was stained with an anti-unknown specificity antibody followed by Alexa Fluor 488 secondary antibody.Flow Cytometry
Flow-cytometry were stained with unimmunized rabbit IgG antibody (black line) or the rabbit IgG-chimeric version of R-125224 (blue line) at a dilution of 1:100 for 1h at RT. After washing, bound antibody was detected using a goat anti-rabbit IgG AlexaFluor 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer. -
Gene Info — FAS
Entrez GeneID
355Protein Accession#
P25445Gene Name
FAS
Gene Alias
ALPS1A, APO-1, APT1, CD95, FAS1, FASTM, TNFRSF6
Gene Description
Fas (TNF receptor superfamily, member 6)
Gene Ontology
HyperlinkGene 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 have been described, some of which are candidates for nonsense-mediated decay (NMD). The isoforms lacking the transmembrane domain may negatively regulate the apoptosis mediated by the full length isoform. [provided by RefSeq
Other Designations
APO-1 cell surface antigen|CD95 antigen|Fas AMA|Fas antigen|OTTHUMP00000020045|OTTHUMP00000020046|OTTHUMP00000020051|OTTHUMP00000059646|apoptosis antigen 1|tumor necrosis factor receptor superfamily member 6|tumor necrosis factor receptor superfamily, mem
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Interactome
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Pathway
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Disease
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