WB (Western Blot)
( Western Blot using anti-Fas antibody R-125224 (Ab00802) Human testis (A) and human ovary (B) lysate samples (35ug protein in RIPA buffer) were resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit IgG version of R-125224 (Ab00802-23.0) 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.)
Fas recombinant antibody
Anti-Fas [R-125224]
Gene Names
FAS; APT1; CD95; FAS1; APO-1; FASTM; ALPS1A; TNFRSF6
Reactivity
Human
Applications
Flow Cytometry, Functional Assay, ELISA
Purity
Protein A Affinity Purified
Synonyms
Fas, Recombinant Antibody; Anti-Fas [R-125224]; h-HFE7A; Tumor necrosis factor receptor superfamily member 6; CD95; CD-95; Apo-1; Apo 1; Apo-1 antigen; Apoptosis-mediating surface antigen FAS; FASLG receptor; Fas recombinant antibody
Reactivity
Human
Clonality
Monoclonal
Isotype
Rabbit IgG, Kappa
Clone Number
R-125224
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.
Purity/Purification
Protein A Affinity Purified
Concentration
1 mg/ml (varies by lot)
Sequence Length
335
Applicable Applications for Fas recombinant antibody
FCM/FACS (Flow Cytometry), ELISA
Immunogen
R-125224 is generated 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 cells.
Buffer Composition
PBS only with 0.02% Proclin 300.
Chimeric Use Statement
This chimeric rabbit antibody was made using the variable domain sequences of the original Human IgG1 format, for improved compatibility with existing reagents, assays and techniques.
Labeled Substance
None
Preparation and Storage
Store at 4 degree C for up to 3 months. For longer storage, aliquot and store at -20 degree C.
WB (Western Blot)
( Western Blot using anti-Fas antibody R-125224 (Ab00802) Human testis (A) and human ovary (B) lysate samples (35ug protein in RIPA buffer) were resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit IgG version of R-125224 (Ab00802-23.0) 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.)
WB (Western Blot)
( Western Blot using anti-Fas antibody R-125224 (Ab00802) Human testis (A) and human ovary (B) lysate samples (35ug protein in RIPA buffer) were resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit IgG version of R-125224 (Ab00802-23.0) 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.)
IF (Immunofluorescence)
( Immunofluorescence staining of fixed MCF7 cells with anti-Fas antibody R-125224 (Ab00802) Immunofluorescence analysis of paraformaldehyde fixed MCF7 cells permeabilized with 0.15% Triton and stained with the chimeric mouse IgG1 version of R-125224 (Ab00802-23.0) 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 Ab00802-23.0, DAPI, merged channels and an isotype control. The isotype control was stained with an anti-unknown specificity antibody (Ab178-23.0) followed by Alexa Fluor® 488 secondary antibody.)
IF (Immunofluorescence)
( Immunofluorescence staining of fixed MCF7 cells with anti-Fas antibody R-125224 (Ab00802) Immunofluorescence analysis of paraformaldehyde fixed MCF7 cells permeabilized with 0.15% Triton and stained with the chimeric mouse IgG1 version of R-125224 (Ab00802-23.0) 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 Ab00802-23.0, DAPI, merged channels and an isotype control. The isotype control was stained with an anti-unknown specificity antibody (Ab178-23.0) followed by Alexa Fluor® 488 secondary antibody.)
FCM/FACS (Flow Cytometry)
( Flow-cytometry using the anti-Fas antibody R-125224 (Ab00802) Jurkat cells were fixed using 2% PFA, permeabilised using 0.5% Triton and stained with unimmunized rabbit IgG antibody (MOPC-21; isotype control, black line) or the rabbit IgG-chimeric version of R-125224 (Ab00802-23.0, 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.)
FCM/FACS (Flow Cytometry)
( Flow-cytometry using the anti-Fas antibody R-125224 (Ab00802) Jurkat cells were fixed using 2% PFA, permeabilised using 0.5% Triton and stained with unimmunized rabbit IgG antibody (MOPC-21; isotype control, black line) or the rabbit IgG-chimeric version of R-125224 (Ab00802-23.0, 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.)
Related Product Information for Fas recombinant antibody
Recombinant monoclonal antibody to Fas. Manufactured using a proprietary Recombinant Platform with variable regions (i.e. specificity) from the hybridoma R-125224.
R-125224 is generated 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 Fas-deficient 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 cells.
R-125224 is generated 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 Fas-deficient 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 cells.
Product Categories/Family for Fas recombinant antibody
References
Haruyama et al. Humanization of the Mouse Anti-Fas Antibody HFE7A and Crystal Structure of the Humanized HFE7A Fab Fragment Biol. Pharm. Bull. 25(12) 1537—1545 (2002) PMID:12499636
NCBI and Uniprot Product Information
NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
11,435 Da
NCBI Official Full Name
tumor necrosis factor receptor superfamily member 6 isoform 1
NCBI Official Synonym Full Names
Fas cell surface death receptor
NCBI Official Symbol
FAS
NCBI Official Synonym Symbols
APT1; CD95; FAS1; APO-1; FASTM; ALPS1A; TNFRSF6
NCBI Protein Information
tumor necrosis factor receptor superfamily member 6
UniProt Protein Name
Tumor necrosis factor receptor superfamily member 6
UniProt Gene Name
FAS
UniProt Synonym Gene Names
APT1; FAS1; TNFRSF6
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Product Notes
The Fas fas (Catalog #AAA72007) is a Recombinant Antibody and is intended for research purposes only. The product is available for immediate purchase. The Anti-Fas [R-125224] reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's Fas can be used in a range of immunoassay formats including, but not limited to, FCM/FACS (Flow Cytometry), ELISA. Researchers should empirically determine the suitability of the Fas fas for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Fas, Monoclonal Recombinant Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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