Mouse Fibronectin ELISA Kit (ab210967)
- Datasheet
- References
- Protocols
Overview
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Product name
Mouse Fibronectin ELISA Kit
See all Fibronectin kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% serum 5 2.3% Inter-assay Sample n Mean SD CV% serum 3 4.4% -
Sample type
Cell culture supernatant, Urine, Serum, Plasma -
Assay type
Sandwich (quantitative) -
Sensitivity
251 pg/ml -
Range
0.63 ng/ml - 40 ng/ml -
Recovery
Sample specific recovery Sample type Average % Range Cell culture supernatant 102 101% - 103% Urine 107 104% - 113% Serum 119 117% - 123% Hep Plasma 108 106% - 109% EDTA Plasma 107 103% - 112% Cit plasma 120 119% - 122% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Mouse
Does not react with: Cow -
Product overview
Fibronectin in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of mouse Fibronectin protein in serum, plasma, urine and cell culture supernatant.
The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
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Notes
Fibronectin is a large glycoprotein present in the extracellular matrix and circulating plasma. Fibronectin is important in many cell adhesion and migration related processes, including wound healing, embryogenesis, nerve regeneration and maintenance of cell shape.
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Tested applications
Suitable for: Sandwich ELISAmore details -
Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Mouse Fibronectin Capture Antibody 1 x 600µl 10X Mouse Fibronectin Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml Antibody Diluent 4BR 1 x 6ml Mouse Fibronectin Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 50ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Research areas
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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
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 localization
Secreted, extracellular space, extracellular matrix. - Information by UniProt
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Alternative names
- CIG
- Cold insoluble globulin
- Cold-insoluble globulin
see all -
Database links
- Entrez Gene: 14268 Mouse
- SwissProt: P11276 Mouse
- Unigene: 193099 Mouse
Associated products
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ELISA kits
Applications
Our Abpromise guarantee covers the use of ab210967 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 |
|---|---|---|
| Sandwich ELISA | Use at an assay dependent concentration. |
Images
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Other - Mouse Fibronectin ELISA Kit (ab210967)SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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Example of mouse Fibronectin standard curve.Background-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of native Fibronectin in mouse serum and plasma samples.The concentrations of Fibronectin were measured in duplicates, interpolated from the Fibronectin standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:4,000, plasma (citrate) 1:4,000, plasma (EDTA) 1: 8,000, and plasma (heparin) 1: 10,000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Fibronectin concentration was determined to be 100 µg/mL in serum, 172 µg/mL in plasma (citrate), 220 µg/mL in plasma (EDTA) and 211 µg/mL in plasma (heparin).
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Interpolated concentrations of native Fibronectin in mouse urine and cell culture supernatant samples.The concentrations of Fibronectin were measured in duplicates, interpolated from the Fibronectin standard curves and corrected for sample dilution. Undiluted samples are as follows: urine 5% and L929 supernatant 10%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Fibronectin concentration was determined to be 979 ng/mL in urine and 300 ng/mL in L929 supernatant.
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Comparison of secreted Fibronectin in unstimulated and PMA/PHA-stimulated L929 CellsL929 cells were grown in the absence (unstimulated) or presence of Phorbol Myristate Acetate (PMA) and phytohemagglutinin (PHA) (stimulated) for 3 days. Fibronectin was measured in 2-fold diluted cell culture supernatants of unstimulated and PMA/PHA stimulated L929 and cell culture media. Measured values were interpolated from the Fibronectin Standard Curve diluted in Sample Diluent NS and corrected for dilution factor. Mean of duplicate values +/-SD are graphed: 300 ng/mL unstimulated, 2,209 ng/mL stimulated, and undetectable in media.
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Comparison of secreted Fibronectin in unstimulated and PMA/PHA-stimulated J774A.1 CellsJ774A.1 cells were grown in the absence (unstimulated) or presence of Phorbol Myristate Acetate (PMA) and phytohemagglutinin (PHA) (stimulated) for 3 days. Fibronectin was measured in 2-fold diluted cell culture supernatants of unstimulated and PMA/PHA stimulated J774A.1 and cell culture media. Measured values were interpolated from the Fibronectin Standard Curve diluted in Sample Diluent NS and corrected for dilution factor. Mean of duplicate values +/-SD are graphed: 294 ng/mL unstimulated, 27.3 ng/mL stimulated, and undetectable in media.
Protocols
References
ab210967 has not yet been referenced specifically in any publications.
