Overview

  • Product name

    Anti-Fibronectin antibody [EPR19241-46]
    See all Fibronectin primary antibodies
  • Description

    Rabbit monoclonal [EPR19241-46] to Fibronectin
  • Host species

    Rabbit
  • Specificity

    Western blot and IHC tests of human samples were negative.

  • Tested applications

    Suitable for: WB, ICC/IF, IP, Flow Cyt, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat
  • Immunogen

    Recombinant fragment within Mouse Fibronectin aa 1350-1650. The exact sequence is proprietary.
    Database link: P11276

  • Positive control

    • WB: Mouse and rat liver lysates; Mouse and rat plasma; Hepa1-6 whole cell lysate. IHC-P: Mouse and rat stomach tissues. ICC/IF: Hepa1-6 cells. Flow Cyt: Hepa1-6 cells. IP: Mouse plasma.
  • 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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab199056 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 1/1000. Detects a band of approximately 273 kDa (predicted molecular weight: 273 kDa).
ICC/IF 1/250.
IP 1/30.
Flow Cyt 1/500.
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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 [EPR19241-46] (ab199056) at 1/1000 dilution

    Lane 1 : Mouse liver lysate
    Lane 2 : Rat liver lysate
    Lane 3 : Mouse plasma
    Lane 4 : Rat plasma
    Lane 5 : Hepa1-6 (mouse hepatoma epithelial cell line) whole cell lysate

    Lysates/proteins at 10 µl/ml buffer per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 273 kDa
    Observed band size: 220-273 kDa
    why is the actual band size different from the predicted?



     

    Exposure time : Lanes 1 and 2:4 seconds; Lanes 3,4 and 5:2 seconds.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression profile and molecular weight observed is consistent with what has been described in the literature (PMID: 6994838, PMID 7516997, PMID: 21923916).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Hepa 1-6 (mouse hepatoma epithelial cell line) cells labeling Fibronectin with ab199056 at 1/250 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and membranous staining on Hepa1-6 cells.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling Fibronectin with ab199056 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on the extracellular matrix of mouse stomach (PMID: 650151).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded rat stomach tissue labeling Fironectin with ab199056 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on the extracellular matrix of rat stomach (PMID: 650151).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized Hepa1-6 (mouse hepatoma epithelial cell line) cell line labeling Fibronectin with ab199056 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

  • Fibronectin was immunoprecipitated from 0.35 mg of mouse plasma with ab199056 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab199056 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: Mouse plasma 10 µg (Input). 

    Lane 2: ab199056 IP in mouse plasma. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab199056 in mouse plasma.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 1 second.

References

This product has been referenced in:

  • Wang XF  et al. DLX5 gene regulates the Notch signaling pathway to promote glomerulosclerosis and interstitial fibrosis in uremic rats. J Cell Physiol 234:21825-21837 (2019). Read more (PubMed: 31297803) »
  • Martínez-Fábregas J  et al. Lysosomal protease deficiency or substrate overload induces an oxidative-stress mediated STAT3-dependent pathway of lysosomal homeostasis. Nat Commun 9:5343 (2018). Read more (PubMed: 30559339) »
See all 2 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 (E8.5 embryos)
Permeabilization
Yes - 0.1% Triton
Specification
E8.5 embryos
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde

Dr. Sophie Astrof

Verified customer

Submitted Oct 10 2019

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (human stomach and colon)
Permeabilization
No
Specification
human stomach and colon
Blocking step
Serum as blocking agent for 15 minute(s) · Concentration: 5µg/mL · Temperature: 22°C
Fixative
peroxidase in methanol

Feifei Song

Verified customer

Submitted Mar 07 2019

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