Anti-Fibronectin antibody (ab299)
- Product nameAnti-Fibronectin antibodySee all Fibronectin primary antibodies ...
- DescriptionRabbit 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 ELISA more details
- Species reactivityReacts with: Human
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
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C (add glycerol to a final volume of 50% for extra stability). Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: None
Constituents: 0.125M Sodium Borate, 0.075M Sodium Chloride, 0.005M EDTA. pH 8.0
- Concentration information loading...
- PurityImmunogen affinity purified
- Purification notesPrepared by immunoaffinity chromatography using immobilized antigens followed by extensive cross-adsorption against human serum proteins and collagen and non-collagen extracellular matrix proteins to remove any unwanted specificities. This product has been sterile filtered.
- Clonality Polyclonal
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.
|ELISA||ELISA: Use at an assay dependent concentration.|
|IP||IP: 1/4000 - 1/8000.|
|WB||WB: 1/4000 - 1/8000.|
|ICC/IF||ICC/IF: Use a concentration of 1 µg/ml.|
|IHC-P||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||sELISA: 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.|
- 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.
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.
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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.
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) »