Overview

  • Product nameAnti-Fibronectin antibody [IST-9]
    See all Fibronectin primary antibodies
  • Description
    Mouse monoclonal [IST-9] to Fibronectin
  • SpecificityThis antibody reacts with an epitope (PEDGIHELFP) located in the ED-A sequence of cellular fibronectin (it therefore only detects the cFN and not extracellular FN (plasma FN)). (Liao et al.) The ED-A segment can be included or omitted from the molecule depending on the pattern of splicing of the mRNA precursors. In transformed or tumour derived cells the ED-A segment is about 10-times higher than in FN from normal fibroblasts, and it may therefore be a significant marker for malignancy (Borsi et al., and others).
  • Tested applicationsWB, RIA, ELISA, IHC-Fr, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Cow, Dog, Human, Pig, Monkey
    Predicted to work with: Rabbit
  • Immunogen

    A region in the ED-A domain of human cellular fibronectin.

  • Positive control
    • normal foetal lung fibroblasts (cell line MRC5), skin, rat or human kidney (see reviews)

Properties

Applications

Our Abpromise guarantee covers the use of ab6328 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
WB 1/400.
RIA Use at an assay dependent dilution.
AP Use at an assay dependent dilution.
ELISA 1/1500.
IHC-Fr Use at an assay dependent dilution.
ICC/IF 1/200.
IHC-P 1/100 - 1/200. Perform enzymatic antigen retrieval before commencing with IHC staining protocol. See Abreviews

Target

  • FunctionFibronectins 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 specificityPlasma 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 diseaseGlomerulopathy with fibronectin deposits 2
  • Sequence similaritiesContains 12 fibronectin type-I domains.
    Contains 2 fibronectin type-II domains.
    Contains 16 fibronectin type-III domains.
  • Developmental stageUgl-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).
    Phosphorylation sites are present in the extracellular medium.
    Proteolytic processing produces the C-terminal NC1 peptide, anastellin.
  • Cellular localizationSecreted > 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 1 antibody
    • FN antibody
    • FN1 antibody
    • FN1 antibody
    • FNZ antibody
    • GFND antibody
    • GFND2 antibody
    • LETS antibody
    • Migration stimulating factor antibody
    • MSF antibody
    • Transformation sensitive protein. antibody
    • Ugl-Y3 antibody
    see all

Anti-Fibronectin antibody [IST-9] images

  • ab6328 staining Fibronectin in Human small intestine tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a Tris-EDTA buffer pH9.0. Samples were incubated with primary antibody (1/50 in blocking buffer) for 30 minutes at 20°C. An undiluted HRP-conjugated Goat anti-mouse  polyclonal was used as the secondary antibody.

    See Abreview

  • This picture shows formalin fixed paraffin embedded rat kidney stained with ab6328 at a dilution of 1:100 after proteinase K digestion.The image was kindly supplied as part of the review submitted by Elizabeth Chlipala.
  • Ab6328 positively staining paraffin fixed canine myocardial infarct. tissue (1/200). The images demonstrate the time course of fibronectin deposition in reperfused canine myocaridal infarcts.
    a: 24 hrs
    b: 7 days
    c: 28 days of reperfusion.

    Secondary: Biotin conjugated goat anti mouse (5 µg/ml). Detection was achieved using DAB.

    This image is an edited version of an image submitted courtesy of an Abreview by Marcin Dobaczewski. We do not have any further information relating to this image.

    See Abreview

  • ab6328 staining human arterial endothelial cells by ICC/IF.  Cells were PFA fixed, permeabilized with 0.05% Triton X-100 and blocked with 0.2% BSA for 1 hour at 25°C  The primary antibody was diluted 1/200 and incubated with the sample for 1 hour at 25°C.  An Alexa Fluor® 488 conjugated donkey anti-mouse antibody, diluted 1/400, was used as the secondary.

    See Abreview

  • Anti-Fibronectin antibody [IST-9] (ab6328) at 1/1000 dilution + whole cell lysate prepared from human platelets ( treated with ADP for 30 minutes) at 20 µg

    Secondary
    HRP conjugated donkey anti-mouse polyclonal at 1/10000 dilution
    developed using the ECL technique

    Exposure time : 2 minutes

    Image courtesy of Dr Mahesh Shivananjappa by Abreview.

    Blocked with 5% milk for 1 hour at 22°C.

    See Abreview

  • All lanes : Anti-Fibronectin antibody [IST-9] (ab6328) at 1/2000 dilution

    Lane 1 : Human dermal fibroblast whole cell lysate, untreated
    Lane 2 : Human dermal fibroblast whole cell lysate, treated with TGF beta 1

    Lysates/proteins at 10 µg per lane.

    Secondary
    HRP-conjugated Goat anti-mouse monoclonal at 1/3000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Exposure time : 15 seconds

    This image is courtesy of an anonymous Abreview

    Blocked with 5% Milk for 1 hour at 20°C.

    See Abreview

References for Anti-Fibronectin antibody [IST-9] (ab6328)

This product has been referenced in:
  • Conway MJ  et al. Mosquito saliva serine protease enhances dissemination of dengue virus into the mammalian host. J Virol 88:164-75 (2014). Read more (PubMed: 24131723) »
  • Ito K  et al. Integrin a9 on lymphatic endothelial cells regulates lymphocyte egress. Proc Natl Acad Sci U S A 111:3080-5 (2014). IHC-P ; Mouse . Read more (PubMed: 24516133) »

See all 48 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 15 minute(s) · Concentration: 1% · Temperature: 20°C
Sample Chinese Hamster Cell (Chinese Hamster Ovary (CHO) cells, CHO-K1)
Specification Chinese Hamster Ovary (CHO) cells, CHO-K1
Permeabilization No
Fixative Paraformaldehyde
Username

Xander van Wijk

Verified customer

Submitted Sep 02 2014

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris-EDTA
Sample Mouse Tissue sections (Liver)
Specification Liver
Permeabilization No
Fixative Formaldehyde
Username

Dr. Suypriya Sen

Verified customer

Submitted Aug 27 2014

The amino acid and converted RNA epitope for ab6328 are given below, along with a link to the converter tool that I used.



Amino acid sequence:

PEDGIHELFP



RNA sequence:

CCCGAAGACGGAAUUCACGAGC...

Read More
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Antigen retrieval step Enzymatic - Buffer/Enzyme Used: pepsin
Sample Rabbit Tissue sections (Liver sections)
Specification Liver sections
Permeabilization No
Fixative Formaldehyde
Username

Ibrahim Badawi

Verified customer

Submitted Dec 07 2013

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Antigen retrieval step Enzymatic - Buffer/Enzyme Used: Pepsin
Sample Rat Tissue sections (Liver sections)
Specification Liver sections
Permeabilization No
Fixative Formaldehyde
Username

Ibrahim Badawi

Verified customer

Submitted Dec 06 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step Serum as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: 25°C
Antigen retrieval step Enzymatic
Sample Rat Tissue sections (GONAD)
Specification GONAD
Permeabilization No
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Sep 23 2013

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 (small intestine)
Specification small intestine
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: TRIS-EDTA Buffer pH 9,0
Permeabilization Yes - Wash Buffer from Dako with Tween
Username

Mr. Rudolf Jung

Verified customer

Submitted Mar 26 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Pig Tissue sections (skin)
Specification skin
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris-EDTA
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Mar 22 2013

Thank you for your generous feedback. I have forwarded this to our team.

I am very pleased to hear you have been provided with satisfactory outcomes in these cases. Please do not hesitate to contact us for any further assitance in the future...

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