Anti-Fibronectin antibody (ab299)
- Datasheet
- References (15)
- Protocols
Overview
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Product nameAnti-Fibronectin antibody
See all Fibronectin primary antibodies -
DescriptionRabbit polyclonal to Fibronectin
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Host speciesRabbit
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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.
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Tested applicationsSuitable for: ELISA, IP, WB, ICC/IF, IHC-P, Sandwich ELISAmore details
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Species reactivityReacts with: Human
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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.
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Positive control
- This antibody gave a positive result in IHC in the following FFPE tissue: Human breast adenocarcinoma
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Add glycerol to a final volume of 50% for extra stability and aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
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Storage bufferpH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 0.87% Sodium chloride, 0.42% Potassium phosphate -
Concentration information loading... -
PurityImmunogen affinity purified
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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.
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Related Products
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. 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
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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.
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Involvement in diseaseGlomerulopathy with fibronectin deposits 2
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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.
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Post-translational
modificationsSulfated.
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 localizationSecreted, extracellular space, extracellular matrix.
- Information by UniProt
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Database links
- Entrez Gene: 2335 Human
- Omim: 135600 Human
- SwissProt: P02751 Human
- Unigene: 203717 Human
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Alternative names
- CIG antibody
- Cold insoluble globulin antibody
- Cold-insoluble globulin antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody (ab299)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.
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Sandwich ELISA - Anti-Fibronectin antibody (ab299)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. -
Immunocytochemistry/ Immunofluorescence - Anti-Fibronectin antibody (ab299)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
This product has been referenced in:
- Li X et al. Upregulation of lactate-inducible snail protein suppresses oncogene-mediated senescence through p16INK4ainactivation. J Exp Clin Cancer Res 37:39 (2018). Read more (PubMed: 29482580) »
- Tanos T et al. Isolation of putative stem cells present in human adult olfactory mucosa. PLoS One 12:e0181151 (2017). Read more (PubMed: 28719644) »
