Overview

  • Product name

    Human Factor IX ELISA Kit, Fluorescent
    See all Factor IX/PTC kits
  • Detection method

    Fluorescent
  • Precision

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

    Serum, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    0.17 ng/ml
  • Range

    0.2 ng/ml - 100 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 108 86% - 130%
    Heparin Plasma 103 85% - 119%
    EDTA Plasma 98 78% - 107%
    Citrate Plasma 104 94% - 121%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat, Rabbit, Goat, Guinea pig, Hamster, Cow, Dog, Pig
  • Product overview

    Factor IX in vitro CatchPoint SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Factor IX protein in humanserum and plasma.


    This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
    If using a Molecular Devices’ plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org.


    The CatchPoint SimpleStep ELISA employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. CatchPoint HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.

  • 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.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    100X Stoplight Red Substrate 1 x 120µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    500X Hydrogen Peroxide (H2O2, 3%) 1 x 50µl
    Antibody Diluent 4BI 1 x 6ml
    Human Factor IX Capture Antibody (Lyophilized) 1 x 600µl
    Human Factor IX Detector Antibody (Lyophilized) 1 x 600µl
    Human Factor IX Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated Black 96-Well Microplate 1 unit
    Stoplight Red Substrate Buffer 1 x 12ml
  • Research areas

  • 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.
  • 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

Applications

Our Abpromise guarantee covers the use of ab229439 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • 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.

     

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

  • 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.

Protocols

References

ab229439 has not yet been referenced specifically in any publications.

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