Overview

  • Product name

    Human Fibrinogen ELISA Kit  (10 x 96 well plate)
    See all Fibrinogen kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 5 3.1%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 3.6%
  • Sample type

    Cell culture supernatant, Saliva, Milk, Urine, Serum, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    188 pg/ml
  • Range

    1.56 ng/ml - 100 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Saliva 113 111% - 117%
    Milk 113 111% - 115%
    Urine 92 89% - 96%
    Serum 99 98% - 99%
    Cell culture media 106 104% - 106%
    Heparin Plasma 108 104% - 110%
    EDTA Plasma 110 104% - 116%
    Citrate Plasma 115 111% - 118%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat, Hamster, Cow, Dog, Pig
  • Product overview

    Fibrinogen in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Fibrinogen protein in human serum, plasma, urine, saliva, milk and cell culture supernatants.


    The 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.  TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    Fibrinogen is a heterohexamer of molecular mass 340 kDa, made up of two sets of alpha, beta, gamma polypeptide chains, and synthesized in the parenchymal cell of the hepatocyte and in the megakaryocyte. Fibrinogen plays a major role in coagulation, and both elevated and decreased levels have clinical significance. Upon cleavage by thrombin in the initial stages of coagulation activation, Fibrinogen self-assembles to yield a fibrin clot matrix that subsequently is crosslinked by factor XIIIa to form an insoluble network. Fibrinogen also binds to the platelet glycoprotein IIb and IIIa receptor so as to form bridges between platelets, thus facilitating aggregation. Elevated plasma Fibrinogen has been identified as an independent risk factor for coronary atherosclerosis and ischemic heart disease. Individuals with congenital absence of Fibrinogen, termed afibrinogenemia, have prolonged bleeding times. Defects in Fibrinogen, alpha are a cause of amyloidosis type 8 (AMYL8) also known as systemic non-neuropathic amyloidosis or Ostertag-type amyloidosis.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 10 x 96 tests
    10X Human Fibrinogen Capture Antibody 1 x 6ml
    10X Human Fibrinogen Detector Antibody 1 x 6ml
    10X Wash Buffer PT 1 x 200ml
    Antibody Diluent CPI 1 x 60ml
    Antibody Diluent CPI 1 x 60ml
    Human Fibrinogen Lyophilized Purified Protein 10 vials
    Plate Seals 10 units
    Sample Diluent NS 2 x 250ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 10 units
    Stop Solution 1 x 120ml
    TMB Development Solution 1 x 120ml
  • Research areas

  • Function

    Fibrinogen has a double function: yielding monomers that polymerize into fibrin and acting as a cofactor in platelet aggregation.
  • Tissue specificity

    Plasma.
  • Involvement in disease

    Defects in FGA are a cause of congenital afibrinogenemia (CAFBN) [MIM:202400]. This is a rare autosomal recessive disorder characterized by bleeding that varies from mild to severe and by complete absence or extremely low levels of plasma and platelet fibrinogen. Note=The majority of cases of afibrinogenemia are due to truncating mutations. Variations in position Arg-35 (the site of cleavage of fibrinopeptide a by thrombin) leads to alpha-dysfibrinogenemias.
    Defects in FGA are a cause of amyloidosis type 8 (AMYL8) [MIM:105200]; also known as systemic non-neuropathic amyloidosis or Ostertag-type amyloidosis. AMYL8 is a hereditary generalized amyloidosis due to deposition of apolipoprotein A1, fibrinogen and lysozyme amyloids. Viscera are particularly affected. There is no involvement of the nervous system. Clinical features include renal amyloidosis resulting in nephrotic syndrome, arterial hypertension, hepatosplenomegaly, cholestasis, petechial skin rash.
  • Sequence similarities

    Contains 1 fibrinogen C-terminal domain.
  • Domain

    A long coiled coil structure formed by 3 polypeptide chains connects the central nodule to the C-terminal domains (distal nodules). The long C-terminal ends of the alpha chains fold back, contributing a fourth strand to the coiled coil structure.
  • Post-translational
    modifications

    The alpha chain is not glycosylated.
    Forms F13A-mediated cross-links between a glutamine and the epsilon-amino group of a lysine residue, forming fibronectin-fibrinogen heteropolymers.
    About one-third of the alpha chains in the molecules in blood were found to be phosphorylated.
    Conversion of fibrinogen to fibrin is triggered by thrombin, which cleaves fibrinopeptides A and B from alpha and beta chains, and thus exposes the N-terminal polymerization sites responsible for the formation of the soft clot. The soft clot is converted into the hard clot by factor XIIIA which catalyzes the epsilon-(gamma-glutamyl)lysine cross-linking between gamma chains (stronger) and between alpha chains (weaker) of different monomers.
    Phosphorylation sites are present in the extracellular medium.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Alternative names

    • FGA
    • FGB
    • FGG
    • Fib2
    • FIBA_HUMAN
    • Fibrinogen A alpha polypeptide
    • Fibrinogen alpha chain
    • Fibrinogen B alpha polypeptide
    • Fibrinogen beta chain
    • Fibrinogen G alpha polypeptide
    • Fibrinogen gamma chain
    • fibrinogen, B beta polypeptide
    • fibrinogen, G gamma polypeptide
    • fibrinogen, gamma polypeptide
    • Fibrinogen--alpha -polypeptide chain
    • Fibrinogen--beta -polypeptide chain
    • Fibrinogen--gamma-polypeptide chain
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab212168 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • ELISA Protocol Summary
  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of Fibrinogen were measured in duplicates, interpolated from the Fibrinogen standard curves and corrected for sample dilution. Undiluted samples are as follows: plasma (citrate) 1:4x104, plasma (EDTA) 1:8x104, and plasma (heparin) 1:4x104. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Fibrinogen concentration was determined to be 1.35 mg/mL in plasma (citrate), 1.45 mg/mL in plasma (EDTA) and 0.62 mg/mL in plasma (heparin).

  • The concentrations of Fibrinogen were measured in duplicates, interpolated from the Fibrinogen standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1.25% and cell culture supernatant 1:800. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Fibrinogen concentration was determined to be 2.8 µg/mL in serum and 10.8 µg/mL in HEPG2 cell culture supernatant.

  • The concentrations of Fibrinogen were measured in duplicates, interpolated from the Fibrinogen standard curves and corrected for sample dilution. Undiluted samples are as follows: urine 12.5%, milk 1.25%, and saliva 1.25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Fibrinogen concentration was determined to be 23.3 ng/mL in urine, 604.9 ng/mL in milk and 718 ng/mL in saliva.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Fibrinogen concentration was determined to be 2.9 µg/mL with a range of 0.88 – 10.5 µg/mL.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Fibrinogen concentration was determined to be 3.9 µg/mL with a range of 1.4 – 6.0 µg/mL.

Protocols

References

ab212168 has not yet been referenced specifically in any publications.

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