Description

Nori Human Fibrinogen ELISA Kit Summary

Alternative names for fibrinogen: Fibrinogen (factor I)

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background: 

Fibrinogen (factor I) is a glycoprotein complex, made in the liver,^[1] that circulates in the blood of all vertebrates.^[2] During tissue and vascular injury, it is converted enzymatically by thrombin to fibrin and then to a fibrin-based blood clot. Fibrin clots function primarily to occlude blood vessels to stop bleeding. Fibrin also binds and reduces the activity of thrombin. This activity, sometimes referred to as antithrombin I, limits clotting.^[1] Fibrin also mediates blood platelet and endothelial cell spreading, tissue fibroblast proliferation, capillary tube formation, and angiogenesis and thereby promotes revascularization and wound healing.^[3] Reduced and/or dysfunctional fibrinogens occur in various congenital and acquired human fibrinogen-related disorders. These disorders represent a group of rare conditions in which individuals may present with severe episodes of pathological bleeding and thrombosis; these conditions are treated by supplementing blood fibrinogen levels and inhibiting blood clotting, respectively.^[4] These disorders may also be the cause of certain liver and kidney diseases.^[1] Fibrinogen is a “positive” acute-phase protein, i.e. its blood levels rise in response to systemic inflammation, tissue injury, and certain other events. It is also elevated in various cancers. Elevated levels of fibrinogen in inflammation as well as cancer and other conditions have been suggested to be the cause of thrombosis and vascular injury that accompanies these conditions.^[5][6] Several disorders in the quantity and/or quality of fibrinogen cause pathological bleeding, pathological blood clotting, and/or the deposition of fibrinogen in the liver, kidneys, and other tissues.

References

1. de Moerloose P, et al. (2013). Seminars in Thrombosis and Hemostasis. 39(6): 585–95.
2. Yong J, Doolittle RF (2003). Proc Natl Acad Sci USA. 100(13): 7527–7532.
3. Mosesson MW (2005). Journal of Thrombosis and Haemostasis. 3(8): 1894–904.
4. Casini A, et al. (2016). Seminars in Thrombosis and Hemostasis. 42(4): 366–74.
5. Davalos D, Akassoglou K (2012). Seminars in Immunopathology. 34(1): 43–62.
6. Repetto O, De Re V (2017). Annals of the New York Academy of Sciences. 1404(1): 27–48.

Product Citations