Overview

  • Product name

    Mouse Fibrinogen ELISA Kit
    See all Fibrinogen kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    EDTA plasma 3 3%
    Inter-assay
    Sample n Mean SD CV%
    EDTA plasma 5 2%
  • Sample type

    Cell culture supernatant, Urine, Serum, Cell culture extracts, Tissue Extracts, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    38 ng/ml
  • Range

    218.75 ng/ml - 14000 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Urine 102 100% - 104%
    Serum 110 101% - 117%
    Tissue Extracts 88 86% - 90%
    Cell culture media 118 112% - 124%
    Heparin Plasma 109 105% - 115%
    EDTA Plasma 106 98% - 111%
    Citrate Plasma 103 98% - 108%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
  • Product overview

    Fibrinogen in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Fibrinogen protein in mouse serum, plasma, urine, cell culture supernatant, and cell and tissue extract samples.


    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.


    Sensitivity:
    Samples in Sample Diluent NS: 38 ng/mL
    Samples in 1X Cell Extraction Buffer PTR: 41 ng/mL

  • 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 hemostasis as one of the primary component of coagulation, and both elevated and decreased levels have clinical significance. The 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. Upon cleavage by thrombin, 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 a fibrinogenemia, 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
    10X Mouse Fibrinogen Capture Antibody 1 x 600µl
    10X Mouse Fibrinogen Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent 5BR 1 x 6ml
    Mouse Fibrinogen Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 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
    • 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 ab213478 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

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

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

  • 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 50%, plasma (EDTA) 1%, plasma (citrate) 10% and plasma (heparin) 1:400. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Fibrinogen concentration was determined to be 6.8 µg/mL in neat serum, 232 µg/mL in neat plasma (EDTA), 146 µg/mL in neat plasma (citrate) and 440 µg/mL in neat 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: 3T3L1 (differentiated, 10 days) 25%, liver (5 days) 100% and lung (6 days) 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Fibrinogen concentration was determined to be 3.1 µg/mL in neat 3T3L1, 3.1 µg/mL in neat liver and 24 µg/mL in neat lung supernatant samples.

  • The concentrations of Fibrinogen were measured in duplicate and interpolated from the Fibrinogen standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Fibrinogen concentration was determined to be 2.2 µg/mL in mouse liver cell tissue extract.

Protocols

References

This product has been referenced in:

  • Tao L  et al. Polypharmacological Profiles Underlying the Antitumor Property of Salvia miltiorrhiza Root (Danshen) Interfering with NOX-Dependent Neutrophil Extracellular Traps. Oxid Med Cell Longev 2018:4908328 (2018). Read more (PubMed: 30210653) »
See 1 Publication for this product

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