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    products/elisa/human-fas-elisa-kit-ab183360.pdf

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Cell Biology Apoptosis Receptors Death Receptors
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RecombinantSimpleStep

Human FAS ELISA Kit (ab183360)

  • Datasheet
  • SDS
  • Protocol Booklet
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SimpleStep ELISA Protocol Diagram
  • Human FAS standard curve comparison
  • Example of human FAS standard curve in Sample Diluent NS.
  • Interpolated concentrations of native FAS in human serum and plasma samples.
  • Interpolated concentrations of native FAS in human PBMC cell culture supernatant derived from cells seeded at 107 cells/mL and cultured in the presence or absence of 1.5% PHA for 36 hours.
  • Serum from ten individual healthy human male donors was measured in duplicate.
  • Sandwich ELISA - Human FAS ELISA Kit (ab183360)

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 26 pg/ml
  • Range: 46.88 pg/ml - 3000 pg/ml
  • Sample type: Cell culture media, Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Serum
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Human

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Overview

  • Product name

    Human FAS ELISA Kit
    See all Fas kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Media 8 3.8%
    Inter-assay
    Sample n Mean SD CV%
    Media 3 9.5%
  • Sample type

    Cell culture supernatant, Serum, Cell culture media, Hep Plasma, EDTA Plasma, Cit plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    26 pg/ml
  • Range

    46.88 pg/ml - 3000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 97 91% - 108%
    Serum 97 88% - 107%
    Cell culture media 100 94% - 105%
    Hep Plasma 105 98% - 111%
    EDTA Plasma 92 81% - 102%
    Cit plasma 98 90% - 105%
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    As of October 15, 2019, Human FAS ELISA kit has been re-developed. We have identified new recombinant monoclonal antibodies to provide improved performance and consistency.

    Human FAS ELISA kit (ab183360) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of FAS protein in human cell culture supernatant, serum, and plasma samples. It uses our proprietary SimpleStep ELISA® technology. Quantitate human FAS with 2 pg/mL sensitivity.

    SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:

            -Single-wash protocol reduces assay time to 90 minutes or less
            -High sensitivity, specificity and reproducibility from superior antibodies
            -Fully validated in biological samples
            -96-wells plate breakable into 12 x 8 wells strips

    A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpeStep ELISA® kits.

  • Notes

    FAS is a plasma membrane receptor for FASLG ligand (TNFSF6/FASLG). Upon ligand binding, the FASLG Receptor is activated and 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) activation mediating apoptosis. FASLG-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 of the FASLG Receptor block apoptosis as they sequester the Fas ligand.

    This kit uses a recombinant protein fragment corresponding to the extracellular portion (amino acid 17-172) of the full length Human Fas Receptor protein as standard sample.

  • Platform

    Microplate

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human FAS Capture Antibody 1 x 600µl
    10X Human FAS Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent CPI - HAMA Blocker (ab193969) 1 x 6ml
    Human FAS Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

    • Cell Biology
    • Apoptosis
    • Receptors
    • Death Receptors
    • Immunology
    • Cell Type Markers
    • CD
    • Cytokines
    • Immunology
    • Innate Immunity
    • Cytokines
    • TNF Superfamily
    • Cell Biology
    • Apoptosis
    • Intracellular
    • Caspases etc
    • Caspases
    • Stem Cells
    • Hematopoietic Progenitors
    • Lymphoid
    • T Lymphocytic Lineage
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Cytokines and cytokine receptors ELISA kits
  • Function

    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).
  • 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.
  • Involvement in disease

    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.
  • Sequence similarities

    Contains 1 death domain.
    Contains 3 TNFR-Cys repeats.
  • Domain

    Contains a death domain involved in the binding of FADD, and maybe to other cytosolic adapter proteins.
  • Cellular localization

    Secreted and Cell membrane.
  • Target information above from: UniProt accession P25445 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • ALPS 1A
    • ALPS1A
    • APO 1
    • Apo 1 antigen
    • APO 1 cell surface antigen
    • Apo-1 antigen
    • APO1
    • Apo1 antigen
    • APO1 cell surface antigen
    • Apoptosis antigen 1
    • Apoptosis mediating surface antigen FAS
    • Apoptosis-mediating surface antigen FAS
    • APT 1
    • APT1
    • CD 95
    • CD 95 antigen
    • CD95
    • CD95 antigen
    • Delta Fas
    • Delta Fas/APO 1/CD95
    • Delta Fas/APO1/CD95
    • Fas
    • Fas (TNF receptor superfamily, member 6)
    • FAS 1
    • FAS 827dupA
    • Fas AMA
    • FAS Antigen
    • Fas cell surface death receptor
    • FAS1
    • FASLG receptor
    • FASTM
    • sFAS
    • Surface antigen APO1
    • TNF receptor superfamily, member 6
    • TNFRSF 6
    • TNFRSF6
    • TNR6_HUMAN
    • Tumor necrosis factor receptor superfamily member 6
    see all
  • Database links

    • Entrez Gene: 355 Human
    • Omim: 134637 Human
    • SwissProt: P25445 Human
    • Unigene: 244139 Human

    Associated products

    • Alternative Versions

      • Human FASLG Receptor Antibody Pair - BSA and Azide free (ab267663)
    • ELISA kits

      • Human Fas ELISA Kit (ab100513)
      • Human FAS ELISA Kit (CD95) (ab119571)

    Images

    • SimpleStep ELISA Protocol Diagram
      SimpleStep ELISA Protocol Diagram

      SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

       

    • Human FAS standard curve comparison
      Human FAS standard curve comparison

      Standard Curve comparison between the original Human FAS SimpleStep ELISA kit and current Human FAS SimpleStep ELISA kit.

    • Example of human FAS standard curve in Sample Diluent NS.
      Example of human FAS standard curve in Sample Diluent NS.

      The FAS standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

    • Interpolated concentrations of native FAS in human serum and plasma samples.
      Interpolated concentrations of native FAS in human serum and plasma samples.

      The concentrations of FAS were measured in duplicates, interpolated from the FAS standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 50%, plasma (citrate, EDTA, heparin) 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean FAS concentration was determined to be 1,393 pg/mL in neat serum, 1,231 pg/mL in neat plasma (citrate) and 1,226 pg/mL in neat plasma (EDTA) and 1,237pg/mL in neat plasma (heparin).

    • Interpolated concentrations of native FAS in human PBMC cell culture supernatant derived from cells seeded at 107 cells/mL and cultured in the presence or absence of 1.5% PHA for 36 hours.
      Interpolated concentrations of native FAS in human PBMC cell culture supernatant derived from cells seeded at 107 cells/mL and cultured in the presence or absence of 1.5% PHA for 36 hours.

      The concentrations of FAS were measured in duplicate and interpolated from the FAS standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean FAS concentration was determined to be 157 pg/mL in neat stimulated PBMC cell culture supernatant and undetectable in unstimulated PBMC cell culture supernatant.

    • Serum from ten individual healthy human male donors was measured in duplicate.
      Serum from ten individual healthy human male donors was measured in duplicate.

      Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean FAS concentration was determined to be 1,513 pg/mL with a range of 1,050 – 2,009 pg/mL.

    • Sandwich ELISA - Human FAS ELISA Kit (ab183360)
      Sandwich ELISA - Human FAS ELISA Kit (ab183360)
      To learn more about the advantages of recombinant antibodies see here.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (0)

    Publishing research using ab183360? Please let us know so that we can cite the reference in this datasheet.

    ab183360 has not yet been referenced specifically in any publications.

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