Overview

  • Product nameAnti-Fibronectin antibodySee all Fibronectin primary antibodies ...
  • Description
    Rabbit polyclonal to Fibronectin
  • SpecificityThis antibody reacts with human Fibronectin and has negligible cross-reactivity with Type I, II, III, IV, V or VI Collagens or Laminin. Minimum Cross Reactivity to collagens and non-collagen extracellular matrix proteins.
  • Tested applicationsELISA, IP, WB, ICC/IF, IHC-P, Sandwich ELISAmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Fibronectin was purified from Human plasma by binding to a denatured gelatin column followed by elution with high concentrations of arginine. The eluted material was further purified by gel filtration.

  • Positive control
    • This antibody gave a positive result in IHC in the following FFPE tissue: Human breast adenocarcinoma

Properties

Applications

Our Abpromise guarantee covers the use of ab299 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
ELISA Use at an assay dependent concentration.
IP 1/4000 - 1/8000.
WB 1/4000 - 1/8000.
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Sandwich ELISA Use a concentration of 0.5 µg/ml. Can be paired for Sandwich ELISA with Mouse monoclonal [HFN 7.1] to Fibronectin - BSA and Azide free (ab80923). For sandwich ELISA, use this antibody as Detection at 0.5 µg/ml with Mouse monoclonal [HFN 7.1] to Fibronectin - BSA and Azide free (ab80923) as Capture.

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 images

  • IHC image of Fibronectin antibody staining in Human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab299, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Standard Curve for Fibronectin (Analyte: Fibronectin protein (Human) (ab81743)); dilution range 1pg/ml to 1µg/ml using Capture Antibody Mouse monoclonal [HFN 7.1] to Fibronectin - BSA and Azide free (ab80923) at 1µg/ml and Detector Antibody Rabbit polyclonal to Fibronectin (ab299) at 0.5µg/ml.
  • ICC/IF image of ab299 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab299, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Protocols

References for Anti-Fibronectin antibody (ab299)

This product has been referenced in:
  • Yoon KD  et al. A novel source of methylglyoxal and glyoxal in retina: implications for age-related macular degeneration. PLoS One 7:e41309 (2012). IP ; Human . Read more (PubMed: 22829938) »
  • Gulhati P  et al. mTORC1 and mTORC2 Regulate EMT, Motility, and Metastasis of Colorectal Cancer via RhoA and Rac1 Signaling Pathways. Cancer Res 71:3246-56 (2011). ICC/IF ; Human . Read more (PubMed: 21430067) »

See all 6 Publications for this product

Product Wall

I found a good set of products that gives a strong sandwich ELISA signal.

Capture: ab80923

Detection: ab299

Antigen: ab81743

Click here (or use the following: http://www.abcam.com/index.html?datasheet=80923)....

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I've included a list of capture, detection, protein, and fluorescent-conjugated secondary below against Fibronectin. The capture and detection antibodies have been tested and guaranteed to work in sandwich ELISA to detect human fibronectin:
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I am very pleased to hear you would like to accept our offer and test ab299 in IHC-P. This code will give you 1 free PRIMARY ANTIBODY before the expi...

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Thank you for your enquiry. We have several antibodies suitable for the labelling of macrophages/monocytes. However, unfortunately many of them have not been tested for theur use in mice. We do have an antibody that recognises the mouse F4/80 antige...

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Thank you for your email. All the information that we have regarding these two antibodies is located on our online datasheets; we do not have any images available at this time. For ab292 I would recommend decreasing the concentration of the primary...

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This antibody has been used in ELISA by using 100uL unlabeled antibody per well at a concentration of 10ug/mL.

This antibody has not been tested in the application or species you describe above, therefore we can't guarantee results. Thesit, triton X100 or Tween 20 are classical solubilizing agents that should work well.

This antibody, used in the applications listed and with the appropriate fluorescent-label-conjugated-secondary antibody, should perform well in immunofluorescence.

The antigen was prepared from Human Plasma. The antibody (immunoblot) shows a 450kDa band non-reduced and 225 kDa band when fully reduced.

Reagents Required: Tris Buffered Saline. Use 10X TBS, pH 7.5 (1.0 M Tris HCl, 1.5 M NaCl) Code # MB-012. Dilute appropriate volume to 1X with deionized water. Store at room temperature up to one month. Lysis Buffer. TBS containing 1.0% of an approp...

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