Abcam, ab108831, Human Factor IX ELISA Kit

Size: 1 x 96Tests
Human Factor IX ELISA Kit is a sandwich ELISA used to quantify Human Factor IX with a sensitivity of 0.50 ng/mL. - Colorimetric sandwich ELISA - 450 nm readout - works on any plate reader - Validated on a number of sample types including cerebrospinal fluid (CSF) - Wide dynamic range – quantifies 1.56 - 100 ng/mL
Key facts
Detection method:Colorimetric,
Sample types:Plasma, Cerebral Spinal Fluid, Serum,
Reacts with:Human,
Assay type:Sandwich (quantitative),
Sensitivity:= 0.5 ng/mL,
Range:1.56 - 100 ng/mL,
Assay time:4h,
Assay Platform:Microplate

Product details:
Human Factor IX ELISA Kit ab108831 is a sandwich ELISA kit to measure Human Factor IX in Serum, Plasma, and Cerebral Spinal Fluid with a sensitivity of 0.50 ng/mL.
How the assay works
Human Factor IX ELISA Kit ab108831 employs a Factor IX specific antibody has been precoated onto 96-well plates and blocked. Standards or test samples are added to the wells and subsequently a Factor IX specific biotinylated detection antibody is added and then followed by washing with wash buffer. StreptavidinPeroxidase Conjugate is added and unbound conjugates are washed away with wash buffer. TMB is then used to visualize StreptavidinPeroxidase enzymatic reaction. TMB is catalyzed by StreptavidinPeroxidase to produce a blue color product that changes into yellow after adding acidic stop solution. The density of yellow coloration is directly proportional to the amount of Factor IX captured in plate.
Assay Specificity
This kit recognizes Factor IX in plasma, serum, and CSF.
This immunoassay is calibrated against a highly purified Human Factor IX. The NIBSC/WHO unclassified purified Human Factor IX preparation 07/182 was evaluated in this kit. The dose response curve of the unclassified standard 07/182 parallels the Factor IX standard curve. To convert sample values obtained with Factor IX Human ELISA Kit (
ab46087
) to approximate NIBSC Units/mL, use the equation below. NIBSC (07/182) approximate value (Units/mL) = 9029.83 x Factor IX value (ng/mL).
Human Factor IX ELISA Kit ab108831 protocol summary:
1. Add samples and standards to wells. Incubate at room temperature.
2. Wash each well and add biotin-antibody conjugate. Incubate at room temperature.
3. Wash each well and add Streptavidin Solution. Incubate at room temperature.
4. Add Chromogen Solution to each well. Incubate at room temperature.
5. Add Stop Solution to each well. Read immediately.
How other researchers are using Human Factor IX ELISA Kit ab108831
Human Factor IX ELISA Kit ab108831 has been used to study targeted overexpression of coagulation factors using CRISPR/Cas9
, the role of Factor IX as a regulator of senescence
, production of coagulation proteins in endothelial cells
, and charactierizing the coagulation cascade in systemic lupus erythematosus
References:
C. Feser, et.al., 2022, PMID: 35563479
P. Carpintero-Fernández, et.al., 2022, PMID: 35184131
C. Cohen, et.al., 2020, PMID: 32029851
Y. Liang, et.al., 2017, PMID: 29599912
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Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions-Multi, Storage information-Please refer to protocols

Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Factor IX also known as Christmas factor or PTC is an important clotting protein in the coagulation cascade. It is a 415 amino acid protein with a mass of approximately 55 kDa. Factor IX is mainly expressed in the liver and then released into the bloodstream where it plays a significant role in blood clotting processes. The recombinant version of this protein referred to as factor IX recombinant is used therapeutically especially in patients who require coagulation support due to factor deficiencies.
Biological function summary
The function of factor IX involves its role in the amplification phase of the coagulation cascade. It participates as part of the intrinsic tenase complex which forms on negatively charged phospholipids in the presence of calcium ions. This complex significantly enhances the conversion of factor X to its active form factor Xa an important step in the clotting process. The activity of factor IX can be assessed using assays such as the factor IX activity assay and ELISA allowing for precise measurement of its function and quantity.
Pathways
Factor IX is integral within the intrinsic pathway of blood coagulation. It interacts with proteins such as factor VIII and factor X. When activated factor IXa forms a complex with the cofactor factor VIIIa on membrane surfaces intensifying the transformation of factor X to factor Xa which then catalyzes the conversion of prothrombin to thrombin. This series of reactions leads to fibrin formation important for stable clot production. Factor IXa also relates to the pathways influencing cellular signaling and hemostasis.
Factor IX deficiencies cause hemophilia B a genetic disorder that leads to improper blood clotting. This disorder is mainly characterized by spontaneous bleeding or severe bleeding after injury. Patients with hemophilia B usually have mutations in the factor IX gene on the X chromosome affecting the protein's function. Successful treatment often involves replacement therapy using factor IX concentrates or the recombinant form. The interaction with proteases such as factor VIIa highlights its importance in therapeutic strategies for bleeding disorders.