Recombinant
RabMAb

Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572)

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Overview

  • Product name
    Anti-Fibrinogen alpha chain antibody [EPR2919]
    See all Fibrinogen alpha chain primary antibodies
  • Description
    Rabbit monoclonal [EPR2919] to Fibrinogen alpha chain
  • Host species
    Rabbit
  • Specificity
    The immunogen is derived from isoform alpha-E, UniProt accession P02671-1, and the antibody is not expected to detect isoform alpha.
  • Tested applications
    Suitable for: Flow Cyt, WB, IP, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    within Human Fibrinogen alpha chain aa 750-850. The exact sequence is proprietary. The immunogen is derived from isoform alpha-E, UniProt accession P02671-1, and the antibody is not expected to detect isoform alpha.
    Database link: P02671-1

  • Positive control
    • Human hepatocellular carcinoma tissue Human plasma, fetal liver and HepG2 whole cell lysate (ab7900) ICC/FC: Hepa1-6 cells WB: Mouse liver cell, mouse plasma cell, mouse platelet cell lysates IHC: Mouse placenta tissue, mouse lung tissue
  • General notes

    Mouse: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab92572 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
Flow Cyt 1/50.
WB 1/1000 - 1/10000. Predicted molecular weight: 95 kDa.
IP 1/50.
IHC-P 1/50 - 1/1000.

Antigen retrieval plus the use of an HRP/AP polymerized secondary antibody is highly recommended for enhanced staining.
ICC/IF 1/250.

Target

  • 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 extracelllular medium.
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Database links
    see all
  • Alternative names
    • Ac1873 antibody
    • Fba5e antibody
    • FGA antibody
    • Fib antibody
    • Fib2 antibody
    • FIBA_HUMAN antibody
    • Fibrinogen alpha chain antibody
    • Fibrinogen, A alpha polypeptide antibody
    • Fibrinogen--alpha polypeptide chain antibody
    • Fibrinopeptide A antibody
    see all

Images

  • Western blot - Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572)
    Western blot - Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572)
    All lanes : Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572) at 1/1000 dilution

    Lane 1 : Mouse liver cell lysate
    Lane 2 : Mouse plasma cell lysate
    Lane 3 : Mouse platelet cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 95 kDa
    Observed band size: 87 kDa
    why is the actual band size different from the predicted?



    The band around 35 kD represents FGA fragment (PMID: 24937656).

    Exposure time:
    Lane 1: 3 minutes;
    Lanes 2-3: 5.5 seconds

     

    Blocking buffer and concentration: 5% NFDM/TBST
    Diluting buffer and concentration: 5% NFDM/TBST

  • Immunohistochemistry paraffin embedded sections - Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572)
    Immunohistochemistry paraffin embedded sections - Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572)

    Immunohistochemical analysis of paraffin-embedded mouse placenta using ab92572 at 1/1000 dilution (0.5 μg/ml). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins. A rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as a secondary antibody (ready to use). Negative control used only the secondary antibody. Counterstained with haematoxylin.

    Positive staining in plasma of mouse placenta.

    The section was incubated with ab92572 for 30 mins at RT. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  • Immunocytochemistry/ Immunofluorescence - Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572)
    Immunocytochemistry/ Immunofluorescence - Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572)

    Immunocytochemistry/ Immunofluorescence analysis of  Hepa1-6 (mouse hepatoma epithelial cell) labeling Fibrinogen alpha chain with ab92572 at a dilution of 1/250. Confocal image showing cytoplasmic staining in Hepa1-6 cells. ab150077 AlexaFluor®488 Goat anti-Rabbit at 1/1000 dilution was used a secondary antibody. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% TritonX-100. ab195889, Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 was used as counterstain and nuclei were stained with DAPI (blue).

     

  • Immunohistochemistry paraffin embedded sections - Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572)
    Immunohistochemistry paraffin embedded sections - Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572)

    Immunohistochemical analysis of paraffin-embedded mouse lung using ab92572 at 1/1000 dilution (0.5 μg/ml). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins, a rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as a secondary antibody (ready to use). Negative control used only the secondary antibody. Counterstained with haematoxylin.

    Strong positive staining in plasma of mouse lung (PMID: 26259584).

    The section was incubated with ab92572 for 30 mins at RT. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  • Flow Cytometry - Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572)
    Flow Cytometry - Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572)
    Flow Cytometry analysis of Hepa1-6 (Mouse hepatoma epithelial cell) labeling Fibrinogen alpha chain with ab92572 at 1/50 (red). Goat anti rabbit IgG (Alexa Fluor® 488), ab150077 at 1/2000 was used as the secondary antibody. Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol.
     
    Isotype control / colour Rabbit monoclonal IgG (ab172730) (black).
    Unlabelled control / colour Cell without incubation with primary antibody and secondary antibody (blue).

     

  • Western blot - Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572)
    Western blot - Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572)
    Rat platelet lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 95 kDa
    Observed band size: 110 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking and diluting buffer: 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572)
    Immunocytochemistry/ Immunofluorescence - Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572)

    Immunocytochemistry/ Immunofluorescence analysis of HepG2 (human hepatocellular carcinoma epithelial cell) labeling Fibrinogen alpha chain with ab92572 at a dilution of 1/250. Confocal image showing cytoplasmic staining in HepG2 cells. ab150077 AlexaFluor®488 Goat anti-Rabbit at 1/1000 dilution was used a secondary antibody. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% TritonX-100. ab195889, Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 was used as counterstain and nuclei were stained with DAPI (blue).

  • Immunoprecipitation - Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572)
    Immunoprecipitation - Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572)

    ab92572 at a dilution of 1/100 immunoprecipitating Fibrinogen alpha chain in human plasma.

    Lane 1 (input): Human plasma (10 µg)
    Lane 2 (+): ab92572 + human plasma.
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab92572 in human plasma.

    For western blotting, ab92572 was used as the primary antibody at a 1/1000 dilution and ab131366 VeriBlot for IP (HRP) was used as the secondary antibody (1/5000).

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572)
    Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using ab92572 at 1/2000 dilution.
  • Western blot - Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572)
    Western blot - Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572)
    All lanes : Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572) at 1/2000 dilution

    Lane 1 : Human plasma lysate
    Lane 2 : Human fetal liver lysate
    Lane 3 : HepG2 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 95 kDa

References

This product has been referenced in:
  • Cheng P  et al. Prevascularization promotes endogenous cell-mediated angiogenesis by upregulating the expression of fibrinogen and connective tissue growth factor in tissue-engineered bone grafts. Stem Cell Res Ther 9:176 (2018). Read more (PubMed: 29973254) »
  • Rancourt RC  et al. Antifibrinolytic mechanisms in acute airway injury after sulfur mustard analog inhalation. Am J Respir Cell Mol Biol 51:559-67 (2014). Read more (PubMed: 24796565) »
See all 2 Publications for this product

Publishing research using ab92572? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Question

Was the immunogen used to make this antibody a 15 or 16 aa peptide corresponding to the sequence of the fibrinopeptide A molecule?

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Answer



The immunogen sequence used to generate ab92572 is a short peptide found with Fibrinogen alpha chain. It does not specifically detect the Fibrinopeptide A molecule.

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Question

Does this product detect the main protein Fibronogen and/or Fibronogen-cleaved proteins? Does this product detect Fibrinopeptide A that has NOT been cleaved from Fibrinogen alpha chain?

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Answer

Thank you for your enquiry. The immunogen contains Fibrinogen alpha chain and is in the Fibrinogen C-terminal region. This antibody detects the main protein Fibrinogen. If Fibrinopeptide A has not been cleaved from Fibrinogen alpha chain, then the antibody is expected to detect Fibrinopeptide A, since it would be part of the existing pro form.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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