Recombinant
RabMAb

Anti-Fibronectin antibody [F1] (ab32419)

Overview

  • Product name
    Anti-Fibronectin antibody [F1]
    See all Fibronectin primary antibodies
  • Description
    Rabbit monoclonal [F1] to Fibronectin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-Fr, WB, IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant full length protein corresponding to Fibronectin aa 1-2400.
    Database link: P02751

  • Positive control
    • WB: Human serum and plasma. SK-OV-3 and HepG2 cell lysate. IHC-P: Human breast carcinoma tissue. ICC/IF: HepG2 and human mesenchymal stem cells. Flow Cytometry: HepG2 cells.
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these 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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab32419 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
IHC-Fr 1/250.
WB 1/1000. Predicted molecular weight: 263 kDa.
IHC-P 1/250 - 1/500.
ICC/IF 1/150.
Flow Cyt 1/20.

Target

  • Function
    Fibronectins bind cell surfaces and various compounds including collagen, fibrin, heparin, DNA, and actin. Fibronectins are involved in cell adhesion, cell motility, opsonization, wound healing, and maintenance of cell shape. Involved in osteoblast compaction through the fibronectin fibrillogenesis cell-mediated matrix assembly process, essential for osteoblast mineralization. Participates in the regulation of type I collagen deposition by osteoblasts.
    Anastellin binds fibronectin and induces fibril formation. This fibronectin polymer, named superfibronectin, exhibits enhanced adhesive properties. Both anastellin and superfibronectin inhibit tumor growth, angiogenesis and metastasis. Anastellin activates p38 MAPK and inhibits lysophospholipid signaling.
  • Tissue specificity
    Plasma FN (soluble dimeric form) is secreted by hepatocytes. Cellular FN (dimeric or cross-linked multimeric forms), made by fibroblasts, epithelial and other cell types, is deposited as fibrils in the extracellular matrix. Ugl-Y1, Ugl-Y2 and Ugl-Y3 are found in urine.
  • Involvement in disease
    Glomerulopathy with fibronectin deposits 2
  • Sequence similarities
    Contains 12 fibronectin type-I domains.
    Contains 2 fibronectin type-II domains.
    Contains 16 fibronectin type-III domains.
  • Developmental stage
    Ugl-Y1, Ugl-Y2 and Ugl-Y3 are present in the urine from 0 to 17 years of age.
  • Post-translational
    modifications
    Sulfated.
    It is not known whether both or only one of Thr-2064 and Thr-2065 are/is glycosylated.
    Forms covalent cross-links mediated by a transglutaminase, such as F13A or TGM2, between a glutamine and the epsilon-amino group of a lysine residue, forming homopolymers and heteropolymers (e.g. fibrinogen-fibronectin, collagen-fibronectin heteropolymers).
    Phosphorylated by FAM20C in the extracellular medium.
    Proteolytic processing produces the C-terminal NC1 peptide, anastellin.
  • Cellular localization
    Secreted, extracellular space, extracellular matrix.
  • Information by UniProt
  • Database links
  • Alternative names
    • CIG antibody
    • Cold insoluble globulin antibody
    • Cold-insoluble globulin antibody
    • DKFZp686F10164 antibody
    • DKFZp686H0342 antibody
    • DKFZp686I1370 antibody
    • DKFZp686O13149 antibody
    • ED B antibody
    • Fibronectin 1 antibody
    • FINC antibody
    • FINC_HUMAN antibody
    • FN antibody
    • FN1 antibody
    • FNZ antibody
    • GFND antibody
    • GFND2 antibody
    • LETS antibody
    • Migration stimulating factor antibody
    • MSF antibody
    • Ugl-Y3 antibody
    see all

Images

  • All lanes : Anti-Fibronectin antibody [F1] (ab32419) at 1/5000 dilution (purified)

    Lane 1 : Human serum
    Lane 2 : Human plasma
    Lane 3 : SK-OV-3 (Human ovarian cancer cell line) cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 263 kDa
    Observed band size: 263 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST

  • Immunohistochemical staining of paraffin embedded human breast carcinoma with purified ab32419 at a dilution of 1/250. The secondary antibody used is ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L), at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • ICC/IF image of unpurified ab32419 stained HepG2 (Human liver hepatocellular carcinoma cell line) cells. The cells were fixed in 4% formaldehyde (10 minutes) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab32419 at 1/100 dilution overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1 hour. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.

  • ICC/IF image of unpurified ab32419 stained human mesenchymal stem cells. The cells were fixed in paraformaldehyde and then incubated in 0.1%BSA / 1% goat serum for 30 minutes, to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32419, 1/100 dilution) for 2 hours at 22°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG. DAPI was used to stain the cell nuclei (blue).

    See Abreview

  • Flow Cytometry analysis of HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling Fibronectin with purified ab32419 at 1/20 dilution (10 µg/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor®488) at 1/2000 dilution was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

  • Anti-Fibronectin antibody [F1] (ab32419) at 1/1000 dilution (purified) + HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate at 20 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 263 kDa
    Observed band size: 263 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST

References

This product has been referenced in:
  • Tang M  et al. Ginsenoside Rg3 inhibits keloid fibroblast proliferation, angiogenesis and collagen synthesis in vitro via the TGF-ß/Smad and ERK signaling pathways. Int J Mol Med 41:1487-1499 (2018). Read more (PubMed: 29328420) »
  • Yu ST  et al. CRLF1 promotes malignant phenotypes of papillary thyroid carcinoma by activating the MAPK/ERK and PI3K/AKT pathways. Cell Death Dis 9:371 (2018). Read more (PubMed: 29515111) »

See all 27 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Skin)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 9
Permeabilization
No
Specification
Skin
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Feb 08 2017

Fibronectin is a highly conserved protein and the full human protein shares 93% homology with cow and 91% homology with rat therefore there is a large chance that an anti-human antibody will also cross react with cow and rat. In addition, there is no s...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Reduced Denaturing (gel 8%)
Sample
Human Cell lysate - whole cell (ovarian cancer cell line: SKOV3)
Specification
ovarian cancer cell line: SKOV3
Blocking step
Blocking one (nacalai tesque) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 27°C
Username

Abcam user community

Verified customer

Submitted Jul 18 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Bone marrow MSCs)
Loading amount
15 µg
Specification
Bone marrow MSCs
Gel Running Conditions
Reduced Denaturing (10% gel)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Sep 11 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Bone marrow MSCs)
Specification
Bone marrow MSCs
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
0.1% BSA + 1% goat serum as blocking agent for 30 minute(s) · Concentration: 1%
Username

Abcam user community

Verified customer

Submitted Sep 11 2012

Vielen Dank für Ihre Anfrage.

Wir haben da mehrere Antikörper, die für Sie in Frage kommen könnten. Ich habe mir erlaubt, zwei Antikörper herauszusuchen, die ich für geeignet halte:

Click here (or use the following: http://www.abc...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (lung, cystic fibrosis)
Specification
lung, cystic fibrosis
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: EDTA, 100C, 20 minutes
Permeabilization
No
Username

Abcam user community

Verified customer

Submitted Dec 02 2009

Thank you for your enquiry. If you refer to form A or B as to alternative splice variants known as ED-A and ED-B (ED stands for extra domain), these alternative splice forms are expressed in a cell-type specific manner. We used plasma fibronectin, w...

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