Overview

  • Product name
  • Description
    Rabbit polyclonal to Fibronectin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ELISA, RIA, ICC/IF, IHC-Fr, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

    Purified fibronectin from murine plasma.

  • Positive control
    • ICC/IF: Normal and cancer-associated fibroblasts; Mouse bone marrow cells. IHC-Fr: Mouse liver tissue. IHC-P: Human aorta tissue; Rat skin tissue. WB: HepG2 cell lysate.
  • General notes

    This antibody may be used to identify fibronectin during wound healing, during tumour progression, and tumour invasion respectively.

Properties

Applications

Our Abpromise guarantee covers the use of ab23750 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.
RIA 1/10000.
ICC/IF 1/40.
IHC-Fr Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 265 kDa (predicted molecular weight: 263 kDa).
IHC-P 1/200 - 1/600. Perform heat mediated antigen retrieval 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

  • Immunofluorescence assay was performed on paired NFs (normal fibroblasts; left) and CAFs (cancer-associated fibroblasts; right) from sample ID 1 using anti-fibronectin (red).

    We isolated CAFs from 15 human lung carcinomas and their corresponding counterpart NFs from their matched non-malignant adjacent tissues, taken at least 10 cm from the outer tumor margin. NFs and CAFs were maintained in 1:1 mixture of DMEM and F12 medium supplemented with 400 ng/ml hydrocortisone, 200 ng/ml insulin, 15% FBS and 1% penicilin/streptomycin. Cells were maintained in 5% CO2 incubator at 37°C.

    For immunofluorescence assay, samples were stained with fibronectin after blocking with bovine serum albumin. Samples were incubated with Alexa Fluor 568-conjugated secondary antibody (red). Microscopic observation was performed under a fluorescence microscope.

  • ab23750 staining Fibronectin in Rat skin tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with Immunoblock® (5 µg/ml) for 30 minutes at 25°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/500 in diluent) for 1 hour 30 minutes at 25°C. An undiluted Goat anti-rabbit HRP/DAB polydetector polyclonal was used as the secondary antibody.

    See Abreview

  • Anti-Fibronectin antibody (ab23750) at 1 µg/ml + HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Predicted band size: 263 kDa
    Observed band size: 265 kDa (why is the actual band size different from the predicted?)

  • ab23750 staining fibronectin in mouse liver tissue by immunohistochemistry (frozen sections). Cells were formaldehyde fixed and permeabilized in 0.2% Triton X-100 prior to blocking in 2% BSA for 30 minutes at 20°C. The primary antibody was diluted 1/200 and incubated with the sample for 9 hours at 4°C. Alexa fluor® 488 goat polyclonal to rabbit Ig, diluted 1/200, was used as the secondary antibody.

    See Abreview

  • ab23750 staining Fibronectin in mouse bone marrow cells by Immunocytochemistry/ Immunofluorescence. Cells were formaldehyde fixed, blocked in 2% BSA for 45 minutes at 20°C. The primary antibody was diluted 1/200 and incubated with sample for 9 hour at 4°C. An Alexa Fluor® 488 conjugated goat polyclonal to rabbit IgG, was used undiluted as secondary. The nuclei were stained with DAPI.

  • ab23750 (2ug/ml) staining Fibronectin in human aorta using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining Fibronectin in the cytoplasm and extracellularly in the connective tissue.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

References

This product has been referenced in:
  • de Bruyn JR  et al. Intestinal fibrosis is associated with lack of response to Infliximab therapy in Crohn's disease. PLoS One 13:e0190999 (2018). Read more (PubMed: 29364909) »
  • Vermeulen M  et al. Development of a Cytocompatible Scaffold from Pig Immature Testicular Tissue Allowing Human Sertoli Cell Attachment, Proliferation and Functionality. Int J Mol Sci 19:N/A (2018). Read more (PubMed: 29329231) »

See all 79 Publications for this product

Customer reviews and Q&As

Filter by Application

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Primary Cardiac Fibroblasts)
Permeabilization
Yes - Triton x-100, 0.01%
Specification
Primary Cardiac Fibroblasts
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde
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Verified customer

Submitted Sep 05 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (prostate)
Total protein in input
100 µg
Immuno-precipitation step
Other - AminoLink Plus Resin
Specification
prostate
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Abcam user community

Verified customer

Submitted Nov 05 2015

Application
Immunocytochemistry
Sample
Mouse Cultured Cells (Fibroblast)
Specification
Fibroblast
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde
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Submitted Oct 26 2015

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse embryonic fibroblast)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
25 µg
Specification
Mouse embryonic fibroblast
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Submitted Sep 30 2015

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Heart)
Permeabilization
Yes - Triton x-100, 0.01%
Specification
Heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde
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Verified customer

Submitted Jun 17 2015

ELISA

Good
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ELISA
Sample
Human Serum (blood)
Specification
blood
Type
Sandwich (Detection)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mrs. Yeri-Alice Rim

Verified customer

Submitted May 23 2013

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (skin)
Specification
skin
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer
Permeabilization
No
Blocking step
immunoblock as blocking agent for 30 minute(s) · Concentration: 5µg/mL · Temperature: 25°C
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Submitted Apr 13 2012

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Medaka fish Tissue sections (Basement membrane)
Specification
Basement membrane
Fixative
Paraformaldehyde
Permeabilization
Yes - acetone
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Username

Dr. Makoto Furutani-Seiki

Verified customer

Submitted Nov 15 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Kidney)
Specification
Kidney
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium Citrate Buffer pH 6.0
Permeabilization
Yes - 0.1% triton X-100
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Verified customer

Submitted Dec 29 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (293FT cells)
Loading amount
100000 cells
Specification
293FT cells
Gel Running Conditions
Reduced Denaturing (12% SDS PAGE)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted May 27 2010

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