Overview

  • Product name
    Anti-Fibronectin antibody [IST-9]
    See all Fibronectin primary antibodies
  • Description
    Mouse monoclonal [IST-9] to Fibronectin
  • Specificity
    This 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 applications
    Suitable for: WB, RIA, ELISA, IHC-Fr, ICC/IF, Other, Blocking, IHC-Pmore details
  • Species reactivity
    Reacts with: Rat, Chicken, Cow, Dog, Human, Pig, Monkey
    Predicted to work with: Mouse, Rabbit
  • Immunogen

    Recombinant fragment corresponding to Human Fibronectin aa 1631-1721. Sequence of the type III repeats termed EIIIA (or ED-A).
    Sequence:

    NIDRPKGLAFTDVDVDSIKIAWESPQGQVSRYRVTYSSPEDGIHELFPAP DGEEDTAELQ GLRPGSEYTVSVVALHDDMESQPLIGTQSTA


    Database link: P02751

  • Epitope
    Liao et al: Clone IST-9, binds to the Ile43 and His44 residues within ED-A in a conformationally dependent fashion, implicating the loop region encompassing both residues as critical for mediating ED-A function. The conformational domain C-C'-E of rat EIIIA encompassing the His44 residue is crucial for constituting the IST-9 epitope.
  • 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 Use a concentration of 1 - 5 µg/ml.
RIA Use at an assay dependent concentration.
AP Use at an assay dependent concentration.
ELISA 1/1500.
IHC-Fr Use at an assay dependent concentration.

Concentration of IST-9 in IHC-fr - depending on the system -  is 0.1 to 2 micrograms per ml.

IST-9 works very well using all iHC-fr procedures.

ICC/IF 1/200.
Other Use at an assay dependent concentration.
Blocking Use at an assay dependent concentration. PubMed: 20643910
IHC-P 1/100 - 1/200. Perform enzymatic antigen retrieval before commencing with IHC staining protocol.

See Abreviews

Concentration of IST-9 in IHC-P is - depending on the system - 2-10 micrograms per ml.

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

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

  • 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

  • ab6328 staining fibronectin in human esophageal epithelial cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with PFA, permeabilized with 1% Triton X-100 in PBS and blocked with 0.25% DAKO serum free protein blocker for 20 minutes at 28°C. Samples were incubated with primary antibody (1/100 in DAKO antibody diluent) for 2 hours at 28°C. ab6785 (1/800) was used as the secondary antibody.

    This photo shows Fibronectin localization in the extracellular matrix of monolayer culture of human esophageal epithelial cells (Het-1A). Nuclei were counterstained with propidium iodide.

    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

  • 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

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

This product has been referenced in:
  • Liu J  et al. Homemade-device-induced negative pressure promotes wound healing more efficiently than VSD-induced positive pressure by regulating inflammation, proliferation and remodeling. Int J Mol Med 39:879-888 (2017). WB ; Rabbit . Read more (PubMed: 28290607) »
  • Bhardwaj A  et al. MYB Promotes Desmoplasia in Pancreatic Cancer through Direct Transcriptional Up-regulation and Cooperative Action of Sonic Hedgehog and Adrenomedullin. J Biol Chem 291:16263-70 (2016). WB ; Human . Read more (PubMed: 27246849) »

See all 67 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Fibroblasts)
Permeabilization
Yes - 0.5% Tritonx100, 10 min
Specification
Fibroblasts
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Apr 07 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rabbit Tissue sections (Iliac vessel)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer
Permeabilization
No
Specification
Iliac vessel
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Feb 27 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (skin)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer
Permeabilization
No
Specification
skin
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jun 10 2016

Application
Western blot
Sample
Mouse Tissue lysate - whole (Cardiac tissue)
Gel Running Conditions
Reduced Denaturing
Loading amount
40 µg
Specification
Cardiac tissue
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Oct 21 2015

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

1-10 of 91 Abreviews or Q&A

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

Sign up