Human Factor IX ELISA Kit, Fluorescent (ab229439)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 0.17 ng/ml
- Range: 0.2 ng/ml - 100 ng/ml
- Sample type: Cit plasma, EDTA Plasma, Hep Plasma, Serum
- Detection method: Fluorescent
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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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, Hep Plasma, EDTA Plasma, Cit 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% 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
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.
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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.
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Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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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 Native Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 50ml SimpleStep Pre-Coated Black 96-Well Microplate 1 unit Stoplight Red Substrate Buffer 1 x 12ml -
Research areas
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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
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. -
Cellular localization
Secreted. - Information by UniProt
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Alternative names
- Christmas Disease
- Christmas factor
- Coagulant factor IX
see all -
Database links
- Entrez Gene: 2158 Human
- Omim: 300746 Human
- SwissProt: P00740 Human
- Unigene: 522798 Human
Images
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Other - Human Factor IX ELISA Kit, Fluorescent (ab229439)
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.
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Example of human Factor IX standard curve in Sample Diluent NSBackground-subtracted data values (mean +/- SD) are graphed.
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Ten individual healthy donors were evaluated for the presence of Factor IX in serum using this assayResults 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.
