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
Product nameHuman Factor IX ELISA Kit
See all Factor IX/PTC kits
Intra-assay Sample n Mean SD CV% Human serum 9 4% Inter-assay Sample n Mean SD CV% Human serum 3 6%
Sample typeSerum, Hep Plasma, EDTA Plasma, Cit plasma
Assay typeSandwich (quantitative)
Range0.78 ng/ml - 50 ng/ml
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 time1h 30m
Assay durationOne step assay
Species reactivityReacts with: Human
Does not react with: Goat, Cow, Pig
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.
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
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.
Storage instructionsStore 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
FunctionFactor 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 specificitySynthesized primarily in the liver and secreted in plasma.
Involvement in diseaseDefects 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 similaritiesBelongs to the peptidase S1 family.
Contains 2 EGF-like domains.
Contains 1 Gla (gamma-carboxy-glutamate) domain.
Contains 1 peptidase S1 domain.
DomainCalcium binds to the gamma-carboxyglutamic acid (Gla) residues and, with stronger affinity, to another site, beyond the Gla domain.
modificationsActivated 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.
- Information by UniProt
- Christmas Disease
- Christmas factor
- Coagulant factor IX
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.
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.
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.
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. 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.
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