Description

Fibrinogen Paired Antibody Set

Affinity’s Fibrinogen Paired Antibody Set consists of matched capture and detecting antibodies only that have been titrated and optimized for use in sandwich style ELISA assays. The product as provided contains sufficient capture and detecting antibodies for five full 96-well microplates and contains a detailed protocol sheet containing directions for use, recipes for solutions and sources for additional materials required. This Fibrinogen Paired Antibody Set is intended to facilitate the end user in establishing an “in-house” immunoassay for research purposes only and must not be used for diagnostic applications.  Assay validation is the responsibility of the end user.

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Product Code: FG-EIA

Supplied Materials:

1. Capture Antibody (FG-EIA-C): One yellow-capped vial containing 0.5 ml of polyclonal affinity purified anti-Fibrinogen antibody for coating plates.
2. Detecting Antibody (FG-EIA-D):  One red-capped tube containing 0.5 ml of peroxidase conjugated affinity-purified polyclonal anti-Fibrinogen antibody for detection of captured Fibrinogen.

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Species Cross Reactivity: View Chart

Product Datasheet:

Fibrinogen Paired Antibody Set for ELISA

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Description of Fibrinogen (Fg)

Human fibrinogen is a 340 kDa plasma protein produced in the liver.  Plasma concentrations are typically 1.7 – 3.5 g/L (5-10 μM).  The intact fibrinogen molecule consists of two identical subunits, each consisting of three non-identical polypeptide chains denoted as Aα, Bβ and γ.  The letters A and B in the Aα and Bβ chains designate, respectively, fibrinopeptide A (FpA, residues 1-16), and fibrinopeptide B (FpB, residues 1-14), which are cleaved by thrombin upon conversion of fibrinogen to fibrin.  The fibrin monomers polymerize in a half-overlap fashion to form insoluble fibrin fibrils. The polymerised fibrin is subsequently stabilized by activated Factor XIII that forms amide linkages between γ chains and, to a lesser extent, α chains of the fibrin molecules.

Proteolysis of fibrinogen by plasmin initially liberates C-terminal residues from the Aα chain to produce fragment X (intact D-E-D, which is still clottable).  Fragment X is further degraded to non-clottable fragments Y (D-E) and D.  Fragment Y can be digested into its constituent D and E fragments.  Proteolysis of crosslinked fibrin by plasmin results in fragment DD (D-Dimer consisting of the D domains of 2 fibrin molecules crosslinked via the γ chains), fragment E (central E domain) as well as DDE in which fragment E is non-covalently associated with DD.  The molecular weights of the cleavage fragments produced from human crosslinked fibrin are: 184 kDa for fragment DD, 92 kDa for D, 50 kDa for E, 1.54 kDa for FpA and 1.57 kDa for FpB.

Most of the fibrinogen in the circulation consists of 2 copies of each chain (Aα₂, Bβ₂, γ_A2), but in normal plasma approximately 10% of the fibrinogen molecules contain one γ_A chain and one variant γ chain (termed γ′), in which the c-terminal AGDV residues are replaced with the amino acid sequence VRPEHPAETEYDSLYPEDDL.  This variant fibrinogen is commonly referred to as fibrinogen gamma prime (γ_A/γ′) but has also been called fibrinogen 2 or peak 2 fibrinogen because it elutes separately from fibrinogen 1 (γ_A2) by ion exchange chromatography.  Residues 414-427 of the γ′ chain of fibrin gamma prime (contain a high-affinity binding site for exosite II of thrombin, which allows the active site of bound thrombin to remain available to interact with substrates while demonstrating resistance to heparin mediated inhibition by antithrombin^1-4.

References and Reviews

1. Hantgan RR, Francis CW, Marder VJ; Fibrinogen Structure and Physiology; in Hemostasis and Thrombosis, 3^rd Edition, eds. RW Colman, J Hirsh, VJ Marder and EW Salzman, pp 277-300, J.B. Lippincott Co., Philadelphia PA, USA, 1994.
2. Binnie CG, Lord ST; The Fibrinogen Sequences that Interact with Thrombin;  Blood 81,  pp 3186-3192, 1993.
3. Pospisil CH, Stafford AR, Fredenburgh JC, Weitz JI; Evidence that both Exosites on Thrombin Participate in Its High Affinity Interaction with Fibrin; JBC 278, pp 21584-21591, 2003.
4. Medved L, Weisel JW; Recommendations for Nomenclature on Fibrinogen and Fibrin; JTH 7, pp 355-359, 2009.

affinity biologicals的凝血–止血ELISA试剂盒VisuLize Antigen试剂盒是凝血-止血ELISA试剂盒系列，用于定量测定VII因子（F7），VIII因子（F8），IX因子（F9），X因子（FX），XI因子（F11）和TAFI抗原使用双抗体酶联免疫吸附试验（ELISA）在人血浆样品中进行检测。