F5 ELISA Kit (Coagulation Factor V/Plasma Factor V)
INTENDED USE
Human F5 ELISA kit can measure concentrations of coagulation factor V (plasma factor V, F5, activated protein C cofactor, PCCF, proaccelerin, labile factor) present is plasma, serum, biological fluids or tissue homogenate samples.
INTRODUCTION
F5, known as coagulation factor V or plasma factor V, is an essential protein for proper blood clotting and hemostasis following vascular injury. As an inactive procofactor, F5 circulates until activated through proteolysis, whereupon it assembles into a calcium ion-dependent complex on cell surfaces to generate thrombin, the central enzyme of blood coagulation. Inherited F5 deficiencies cause a bleeding disorder, while mutation-induced hyperactivity increases thrombotic risk.
Produced in liver and megakaryocytes then entering circulation in procofactor form, intact F5 requires proteolytic activation by thrombin or Factor Xa, which splits F5 into active fragments F5a and F5b. These fragmented subunits then come together into a calcium-stabilized complex on platelet membrane surfaces to substantially boost thrombin production. This amplifying feedback loop localizing thrombin generation to platelet membranes prevents unwarranted clotting. The F5 protein complex acts as a necessary cofactor for timely thrombin catalytic function, which converts fibrinogen into fibrin strands forming vessel-occluding blood clots. Natural anticoagulants like Protein C eventually halt clot growth by degrading F5a and limiting further thrombin productivity.
While F5 activity facilitates beneficial hemostatic plug formation following vascular injury, dysfunction promoting either insufficient or excessive F5 coagulation responses can cause bleeding or thrombosis issues. F5 deficiency, typically inherited rather than acquired, leads to rare bleeding disorders stemming from inadequate thrombin generation. Much more common are thrombophilic F5 gene variants causing resistance to Protein C regulation, enabling amplified thrombin production and recurring venous thromboembolism (VTE).
CONTENT
All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.
- One 96-Well Plate: Pre-coated with anti- F5.
- Standards: Lyophilized recombinant.
- Sample/Standard Dilution Buffer.
- Biotinylated-labelled Antibody.
- Antibody Dilution Buffer.
- HRP-Streptavidin Conjugate (SABC).
- SABC Dilution Buffer.
- TMB Substrate.
- Wash Buffer (25x).
- Plate Sealer.
- Product Instructions.
TYPICAL RESULTS
For this F5 ELISA kit, it is recommended that a standard curve is generated for each assay carried out.
Standard Curve: 0, 15.625, 31.25, 62.5, 125, 250, 500, 1000 pg/ml.
Reactivity: Human
Sensitivity: 9.375 pg/ml
Range: 15.625 – 1000 pg/ml
Principle: Sandwich, Double Antibody
Application: Research Use Only.
ASSAY CHARACTERISTICS
– Specificity: Highly specific for F5, no cross reactivity or interference between F5 and analogues was detected.
– Recovery: Serum (91 – 100%), EDTA Plasma (86 – 102%), Heparin Plasma (86 – 103%).
– Linearity: Serum (94 – 105%), EDTA Plasma (90 – 100%), Heparin Plasma (80 – 87%).
– Precison Intra-Assay: CV < 8%
– Precison Inter-Assay: CV < 10%
REFERENCES
- Coagulation factor V and thrombophilia: background and mechanisms. Thromb Haemost. (2007) 98 (3): 530-42. Segers K., et al.
- Activated factor V is a cofactor for the activation of factor XI by thrombin in plasma. Proc Natl Acad Sci U S A. (2010) 107 (20): 9083-7. Maas C., et al.
- Factor V Inhibitors: A Diagnostic and Therapeutic Challenge. Arch Pathol Lab Med. (2017) 141 (12): 1728-1731. Olson N.J. and Ornstein D.L.
- A new look at blood coagulation factor V. Curr Opin Hematol. (2011) 18 (5): 338-42. Camire R.M.
ADDITIONAL INFORMATION
- Full Name: F5 ELISA Kit (Coagulation Factor V/Plasma Factor V)
- Reactivity: Human
- Sample Type: Plasma, Tissue Homogenates, Serum, Biological Fluids
- Sensitivity: 9.375 pg/ml
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