Overview

  • Product name
    Anti-Fibronectin antibody [F14]
    See all Fibronectin primary antibodies
  • Description
    Rabbit monoclonal [F14] to Fibronectin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-Fr, WB, ICC/IF, Flow Cytmore details
    Unsuitable for: IHC-P
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

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

  • Positive control
    • WB: Human, mouse and rat serum lysate. IHC-Fr: Rat kidney tissue. IHC-P: Human uterus tissue. ICC/IF: HepG2 cells.
  • General notes

    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 ab45688 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 Use at an assay dependent concentration.
WB 1/1000 - 1/10000. Detects a band of approximately 263 kDa (predicted molecular weight: 263 kDa).
ICC/IF 1/500.

For unpurified use at 1/250.

Flow Cyt 1/150.
  • Application notes
    Is unsuitable for IHC-P.
  • 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

    • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
      Lane 2: Fibronectin knockout HAP1 whole cell lysate (20 µg)

      Lanes 1 - 2: Merged signal (red and green). Green - ab45688 observed at 262 kDa. Red - loading control, ab18058, observed at 130 kDa.

      ab45688 was shown to react with Fibronectin in wild-type HAP1 cells as signal was lost in Fibronectin knockout cells. Wild-type and Fibronectin knockout samples were subjected to SDS-PAGE. Ab45688 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/500 dilution and 1/2000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling Fibronectin with purified ab45688 at 1:150 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

    • Immunocytochemistry/ Immunofluorescence analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling Fibronectin with Purified ab45688 at 1:500 dilution. Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200. ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • Anti-Fibronectin antibody [F14] (ab45688) at 1/10000 dilution (purified) + Human serum lysates at 15 µg

      Secondary
      Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

      Predicted band size: 263 kDa



      Blocking and diluting buffer: 5% NFDM/TBST.

    • Unpurified ab45688 staining Fibronectin in rat kidney tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde, permeabilized with 0.1% Triton and blocked with Cas-Block for 30 minutes at 25°C. Samples were incubated with primary antibody (1/250) for 16 hours at 4°C. An Alexa Fluor®488-conjugated Goat anti-rabbit IgG polyclonal (1/400) was used as the secondary antibody.

      See Abreview

    • All lanes : Anti-Fibronectin antibody [F14] (ab45688) at 1/10000 dilution (purified)

      Lane 1 : Mouse serum lysates
      Lane 2 : Rat serum lysates

      Lysates/proteins at 15 µg per lane.

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

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



      Blocking and diluting buffer: 5% NFDM/TBST

    • Anti-Fibronectin antibody [F14] (ab45688) at 1/5000 dilution (unpurified) + human serum

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

    • Paraffin-embedded human uterus tissue stained for Fibronectin with unpurified ab45688 at a 1/250 dilution in immunohistochemical analysis.

    • ICC/IF image of unpurified ab45688 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 ab45688 at 1/250 dilution overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1 hour. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43 µM for 1 hour at room temperature.

    References

    This product has been referenced in:
    • Han Y  et al. Reactive oxygen species promote tubular injury in diabetic nephropathy: The role of the mitochondrial ros-txnip-nlrp3 biological axis. Redox Biol 16:32-46 (2018). Read more (PubMed: 29475133) »
    • Ng PK  et al. Systematic Functional Annotation of Somatic Mutations in Cancer. Cancer Cell 33:450-462.e10 (2018). Read more (PubMed: 29533785) »
    See all 17 Publications for this product

    Customer reviews and Q&As

    1-2 of 2 Abreviews or Q&A

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Kidney)
    Permeabilization
    Yes - 0.1% TritonX-100
    Specification
    Kidney
    Blocking step
    Cas-Block (Invitrogen) as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: RT°C
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Jun 13 2017

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Rat Tissue sections (Kidney)
    Permeabilization
    Yes - 0.1% TritonX-100
    Specification
    Kidney
    Blocking step
    Cas-Block (Invitrogen) as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: RT°C
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Jun 13 2017

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

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