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    products/elisa/human-factor-ix-elisa-kit-ab188393.pdf

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SimpleStep

Human Factor IX ELISA Kit (ab188393)

  • Datasheet
  • SDS
  • Protocol Booklet
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Other - Human Factor IX ELISA Kit (ab188393)
  • Example of Factor IX standard curve in Sample Diluent NS.
  • Example of Factor IX standard curve in Sample Diluent NS.
  • Linearity of dilution.
  • Ten individual healthy donors were evaluated for the presence of Factor IX in serum using this assay.
  • Mouse plasma hFIX protein levels as measure by ELISA

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 0.23 ng/ml
  • Range: 0.78 ng/ml - 50 ng/ml
  • Sample type: 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 Factor IX ELISA Kit
    See all Factor IX/PTC kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Human serum 9 4%
    Inter-assay
    Sample n Mean SD CV%
    Human serum 3 6%
  • Sample type

    Serum, Hep Plasma, EDTA Plasma, Cit plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    0.23 ng/ml
  • Range

    0.78 ng/ml - 50 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 108 86% - 130%
    Hep Plasma 103 85% - 119%
    EDTA Plasma 98 78% - 107%
    Cit plasma 104 94% - 121%
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
    Does not react with: Goat, Cow, Pig
  • Product overview

    Human Factor IX ELISA Kit (ab188393) has been re-developed. We have identified new recombinant monoclonal antibodies to provide improved performance and consistency. This kit will be discontinued when stock is depleted. The new version of the kit is available as ab300307.


    Human Factor IX ELISA Kit (ab188393) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Factor IX protein in cit plasma, edta plasma, hep plasma, and serum. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human Factor IX with 0.23 ng/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 SimpleStep ELISA® kits.


    SPECIES REACTIVITY


    This kit recognizes human Factor IX protein.


    Other species reactivity was determined by measuring 1% serum samples of various species, interpolating the protein concentrations from the human standard curve, and expressing the interpolated concentrations as a percentage of the protein concentration in human serum assayed at the same dilution.


    Reactivity < 3% was determined for the following species: Mouse, Rat, Hamster, Guinea Pig , Rabbit, Dog, Goat, Pig, Cow

  • Notes

    Factor IX, also known as Christmas Factor, is a 52 kDa zymogen of plasma serine proteases that initiates coagulation through activation by Tissue Factor (TF) and Factor VIIa. The activated form of Factor IX, known as Factor IXa, further activates an intrinsic activation complex that consists of factor X and factor VIIIa in the presence of Ca (2+) ions, and phospholipids. Mutations that cause a deficiency in Factor IX lead to x-linked hemophilia B, whereas overproduction of Factor IX has been shown to be a risk factor in venous thromboembolism (VTE). In 2013 the Food and Drug Administration approved the use of recombinant factor IX as a prophylactic treatment of patients with hemophilia B.

  • Platform

    Microplate

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 4BI 1 x 6ml
    Human Factor IX Capture Antibody (Lyophilized) 2 vials
    Human Factor IX Detector Antibody (Lyophilized) 2 vials
    Human Factor IX Lyophilized Native 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

    • Cardiovascular
    • Blood
    • Other
    • Cardiovascular
    • Blood
    • Coagulation
    • Intrinsic
    • Immunology
    • Innate Immunity
    • Complement
    • Classical Pathway
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Blood coagulation ELISA kits
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Cardiovascular ELISA kits
  • Function

    Factor IX is a vitamin K-dependent plasma protein that participates in the intrinsic pathway of blood coagulation by converting factor X to its active form in the presence of Ca(2+) ions, phospholipids, and factor VIIIa.
  • Tissue specificity

    Synthesized primarily in the liver and secreted in plasma.
  • Involvement in disease

    Defects in F9 are the cause of recessive X-linked hemophilia B (HEMB) [MIM:306900]; also known as Christmas disease.
    Note=Mutations in position 43 (Oxford-3, San Dimas) and 46 (Cambridge) prevents cleavage of the propeptide, mutation in position 93 (Alabama) probably fails to bind to cell membranes, mutation in position 191 (Chapel-Hill) or in position 226 (Nagoya OR Hilo) prevent cleavage of the activation peptide.
    Defects in F9 are the cause of thrombophilia due to factor IX defect (THR-FIX) [MIM:300807]. A hemostatic disorder characterized by a tendency to thrombosis.
  • Sequence similarities

    Belongs to the peptidase S1 family.
    Contains 2 EGF-like domains.
    Contains 1 Gla (gamma-carboxy-glutamate) domain.
    Contains 1 peptidase S1 domain.
  • Domain

    Calcium binds to the gamma-carboxyglutamic acid (Gla) residues and, with stronger affinity, to another site, beyond the Gla domain.
  • Post-translational
    modifications

    Activated by factor XIa, which excises the activation peptide.
    The iron and 2-oxoglutarate dependent 3-hydroxylation of aspartate and asparagine is (R) stereospecific within EGF domains.
  • Cellular localization

    Secreted.
  • Target information above from: UniProt accession P00740 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • Christmas Disease
    • Christmas factor
    • Coagulant factor IX
    • Coagulation factor 9
    • Coagulation factor IX
    • Coagulation factor IXa heavy chain
    • F9
    • FA9_HUMAN
    • Factor 9
    • Factor IX Deficiency
    • FactorIX
    • FIX
    • Haemophilia B
    • HEMB
    • MGC129641
    • MGC129642
    • P19
    • Plasma Thromboplastic Component
    • Plasma thromboplastin component
    • PTC
    see all
  • Database links

    • Entrez Gene: 2158 Human
    • Omim: 300746 Human
    • SwissProt: P00740 Human
    • Unigene: 522798 Human

    Associated products

    • ELISA kits

      • Human Factor IX ELISA Kit (ab108831)
      • Human Factor IX ELISA Kit (with plasma controls) (ab168546)

    Images

    • Other - Human Factor IX ELISA Kit (ab188393)
      Other - Human Factor IX ELISA Kit (ab188393)

      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.

       

    • Example of Factor IX standard curve in Sample Diluent NS.
      Example of Factor IX standard curve in Sample Diluent NS.

      Background-subtracted data values (mean +/- SD) are graphed.

    • Example of Factor IX standard curve in Sample Diluent NS.
      Example of Factor IX standard curve in Sample Diluent NS.

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

    • Linearity of dilution.
      Linearity of dilution.

      Native Factor IX was measured in the following biological samples in a 2-fold dilution series. Sample dilutions are made in Sample Diluent NS.

      Linearity of dilution is determined based on interpolated values from the standard curve. Linearity of dilution defines a sample concentration interval in which interpolated target concentrations are directly proportional to sample dilution.

    • Ten individual healthy donors were evaluated for the presence of Factor IX in serum using this assay.
      Ten individual healthy donors were evaluated for the presence of Factor IX in serum using this assay.

      Results were interpolated from the standard curve in Sample Diluent NS and corrected by sample dilution (400 fold). The mean level of Factor IX is 1,240 ng/mL with a range of 246 – 3,994 ng/mL and a standard deviation of 1,193 ng/mL.

    • Mouse plasma hFIX protein levels as measure by ELISA
      Mouse plasma hFIX protein levels as measure by ELISAImage from Lulu Haung et al, Genome Biology, 19, 4, figure 5; doi: 10.1186/s13059-017-1386-9

      Mouse plasma hFIX protein levels as measure by ELISA. Statistical significance was determined using a one-way ANOVA and Dunnett’s multiple comparison test in Prism. All groups were compared to DPBS-treated hFIX-R29X group. ** p < 0.01; *** p < 0.001; **** p < 0.0001.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (5)

    Publishing research using ab188393? Please let us know so that we can cite the reference in this datasheet.

    ab188393 has been referenced in 5 publications.

    • De Giorgi M  et al. Targeting the Apoa1 locus for liver-directed gene therapy. Mol Ther Methods Clin Dev 21:656-669 (2021). PubMed: 34141821
    • Cohen CT  et al. Production and control of coagulation proteins for factor X activation in human endothelial cells and fibroblasts. Sci Rep 10:2005 (2020). PubMed: 32029851
    • Chen J  et al. CRISPR/Cas9-mediated knockin of human factor IX into swine factor IX locus effectively alleviates bleeding in hemophilia B pigs. Haematologica N/A:N/A (2020). PubMed: 31974191
    • Huang L  et al. Targeting Translation Termination Machinery with Antisense Oligonucleotides for Diseases Caused by Nonsense Mutations. Nucleic Acid Ther 29:175-186 (2019). PubMed: 31070517
    • Huang L  et al. Antisense suppression of the nonsense mediated decay factor Upf3b as a potential treatment for diseases caused by nonsense mutations. Genome Biol 19:4 (2018). PubMed: 29334995

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