Overview

  • Product name
    Human Fibrinogen ELISA Kit, Fluorescent
    See all Fibrinogen kits
  • Detection method
    Fluorescent
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    serum 8 3.4%
    Inter-assay
    Sample n Mean SD CV%
    serum 3 11.7%
  • Sample type
    Cell culture supernatant, Saliva, Milk, Urine, Serum, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    29 pg/ml
  • Range
    15.63 pg/ml - 64000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 94 93% - 95%
    Saliva 101 98% - 104%
    Milk 103 102% - 105%
    Urine 100 99% - 100%
    Serum 91 90% - 94%
    Heparin Plasma 95 95% - %
    EDTA Plasma 93 91% - 94%
    Citrate Plasma 94 91% - 96%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Mouse, Human
    Does not react with: Rat
  • Product overview

    Fibrinogen in vitro CatchPoint 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 supernatant samples.


    This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
    If using a Molecular Devices’ plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org.


    The CatchPoint 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.  CatchPoint HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.


    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 1 x 96 tests
    100X Stoplight Red Substrate 1 x 120µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    500X Hydrogen Peroxide (H2O2, 3%) 1 x 50µl
    Antibody Diluent 4BI 1 x 6ml
    10X Human Fibrinogen Capture Antibody 1 x 600µl
    10X Human Fibrinogen Detector Antibody 1 x 600µl
    Human Fibrinogen Lyophilized Purified Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated Black 96-Well Microplate 1 unit
    Stoplight Red Substrate Buffer 1 x 12ml
  • 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
    • 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 ab229419 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
  • The Fibrinogen standard curve was prepared as described in Section 10. Raw data generated on SpectraMax M4 Multi-Mode Microplate Reader is shown in the table. 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: serum 1:670, plasma (citrate) 1:5x105, plasma (EDTA) 1:5x105, and plasma (heparin) 1:5x105. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Fibrinogen concentration was determined to be 3.45 µg/mL in serum, 2.86 mg/mL in plasma (citrate), 2.96 mg/mL in plasma (EDTA), and 2.64 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: urine 1:1.7, saliva 1:133, milk 1:133 and HepG2 supernatant 1:500. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Fibrinogen concentration was determined to be 5.93 ng/mL in urine, 934.5 ng/mL in saliva, 199.7 ng/mL in milk and 1.35 µg/mL in HepG2 supernatant.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Fibrinogen concentration was determined to be 1,070 ng/mL with a range of 647 – 2,027 ng/mL.

Protocols

References

ab229419 has not yet been referenced specifically in any publications.

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