Human Fibronectin ELISA Kit (ab219046)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 20.6 pg/ml
- Range: 125 pg/ml - 8000 pg/ml
- Sample type: Cell culture extracts, Cell culture supernatant, Cit plasma, EDTA Plasma, Milk, Serum, Tissue Extracts
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human, Rhesus monkey
Overview
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Product name
Human Fibronectin ELISA Kit
See all Fibronectin kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Serum 3 5% Inter-assay Sample n Mean SD CV% Serum 5 8.3% -
Sample type
Cell culture supernatant, Milk, Serum, Cell culture extracts, Tissue Extracts, EDTA Plasma, Cit plasma -
Assay type
Sandwich (quantitative) -
Sensitivity
20.6 pg/ml -
Range
125 pg/ml - 8000 pg/ml -
Recovery
Sample specific recovery Sample type Average % Range Cell culture supernatant 102 90% - 117% Milk 102 91% - 111% Serum 93 87% - 102% Cell culture extracts 89 80% - 105% Tissue Extracts 90 80% - 131% EDTA Plasma 86 81% - 94% Cit plasma 99 87% - 120% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human, Rhesus monkey
Does not react with: Cow -
Product overview
Human Fibronectin ELISA Kit (ab219046) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Fibronectin protein in cell culture supernatant, cit plasma, edta plasma, milk, serum, tissue extracts, and cell culture extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human Fibronectin with 20.6 pg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells stripsA 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
ASSAY SPECIFICITY This kit recognizes both native and recombinant human Fibronectin protein in serum, plasma (citrate and EDTA), milk, cell culture supernatant, and cell and tissue extract samples only.
CROSS REACTIVITY Recombinant human Anastelin was prepared within the working range of the assay and assayed for cross reactivity. No cross-reactivity was observed.
SPECIES REACTIVITY This kit recognizes human Fibronectin protein.Other species reactivity was determined by measuring 1/100,000 serum (mouse, rat, cow) or plasma (Rhesus macaque) samples of various species, interpolating the protein concentrations from the human standard curve, and expressing the interpolated concentrations as a percentage of the protein concentration in human serum or plasma assayed at the same dilution.
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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 and nerve regeneration. Differential expression of fibronectin is seen in coronary heart disease, glomerulopathy and tumor cell metastasis. The protein contains binding sites for collagen, heparin and fibrin and is a specific ligand for several integrin adhesion receptors. Fibronectin exists as a dimer or multimer.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
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 Wash Buffer PT (ab206977) 1 x 20ml 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent 4BR 1 x 6ml 10X Human Fibronectin Capture Antibody 1 x 600µl 10X Human Fibronectin Detector Antibody 1 x 600µl Human 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: 2335 Human
- Omim: 135600 Human
- SwissProt: P02751 Human
- Unigene: 203717 Human
Images
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Other - Human Fibronectin ELISA Kit (ab219046)
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 human Fibronectin standard curve in Sample Diluent NS.Background-subtracted data values (mean +/- SD) are graphed.
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Example of human Fibronectin standard curve in Sample Diluent NS.Example of human Fibronectin standard curve in Sample Diluent NS. The Fibronectin standard curve was prepared as described. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of native Fibronectin in human 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 1/20,000, plasma (citrate) 1/20,000, and plasma (EDTA) 1/40,000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Fibronectin concentration was determined to be 126.5 µg/mL in serum, 141.5 µg/mL in plasma (citrate), and 232.1 µg/mL in plasma (EDTA).
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Example of human Fibronectin standard curve in 1X Cell Extraction Buffer PTR.Background-subtracted data values (mean +/- SD) are graphed.
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Example of human Fibronectin standard curve in 1X Cell Extraction Buffer PTR.Example of human Fibronectin standard curve in 1X Cell Extraction Buffer PTR. The Fibronectin standard curve was prepared as described. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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Human Fibronectin standard curve comparison dataStandard curve comparison between human Fibronectin SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows comparable sensitivity.
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Interpolated concentrations of native Fibronectin in human breast milk and HepG2 cell culture supernatant (4 days) 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: milk 1/100, and HepG2 cell culture supernatant 1/2000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Fibronectin concentration was determined to be 502.9 ng/mL in milk, and 9683 ng/mL in HepG2 cell culture supernatant.
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Interpolated concentrations of native Fibronectin in HepG2, SW 480 and U87-MG cell extracts.Interpolated concentrations of native Fibronectin in human HepG2 cell extract samples based on a 25 μg/mL extract load, SW 480 cell extract samples based on a 100 μg/mL extract load, and U87-MG cell extract samples based on a 5 μg/mL extract load. The concentrations of Fibronectin were measured in duplicate and interpolated from the Fibronectin standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Fibronectin concentration was determined to be 7085 pg/mL in HepG2 cell extract, 6314 pg/mL in SW 480 cell extract, and 1749 pg/mL in U87-MG cell extract.
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Interpolated concentrations of native Fibronectin in various tissue extracts.Interpolated concentrations of native Fibronectin in human liver tissue extract based on a 100 μg/mL extract load, heart tissue extract based on a 100 μg/mL extract load, skeletal muscle based on a 100 μg/mL extract load, colon tissue extract based on a 5 μg/mL extract load, and placenta tissue extract based on a 1.25 μg/mL extract load. The concentrations of Fibronectin were measured in duplicate and interpolated from the Fibronectin standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Fibronectin concentration was determined to be 7634 pg/mL in liver tissue extract, 4910 pg/mL in heart tissue extract, 4278 pg/mL in skeletal muscle tissue extract, 2684 pg/mL in colon tissue extract, and 1224 pg/mL in placenta tissue extract.
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Serum from ten individual healthy male human donors was measured in duplicate.Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Fibronectin concentration was determined to be 146.2 µg/mL with a range of 35.99 – 319.2 µg/mL.
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Assay sensitivity.The MDD was determined by calculating the mean of zero standard replicates and adding 2 standard deviations then extrapolating the corresponding concentration.
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Interpolated concentrations of native Fibronectin in human cell and tissue extract samples.The concentrations of Fibronectin were measured in three different dilutions in duplicate and interpolated from the Fibronectin standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted in ng of Fibronectin per mg of extract (mean +/- SD, n=3). Fibronectin concentration was determined to be 271.8 ng/mg in HepG2 cell extract, 65.52 ng/mg in SW 480 cell extract, 359.6 ng/mg in U87-MG cell extract, 75.22 ng/mg in liver tissue extract, 47.20 ng/mg in heart tissue extract, 43.52 ng/mg in skeletal muscle tissue extract, 557.9 ng/mg in colon tissue extract, and 998.8 ng/mg in placenta tissue extract samples.
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Linearity of dilution.Linearity of dilution is determined based on interpolated values from the standard curve. Linearity of dilution defines a sample concentration interval in which interpolated target concentrations are directly proportional to sample dilution.
Native Fibronectin was measured in serum, plasma, de-fatted breast milk, and cell culture supernatant (SN) samples in a 2-fold dilution series. Sample dilutions are made in Sample Diluent NS.
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Linearity of dilution.Native Fibronectin was measured in the following biological samples in a 2-fold dilution series. Sample dilutions are made in 1X Cell Extraction Buffer PTR.
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Linearity of dilution.Native Fibronectin was measured in the following biological samples in a 2-fold dilution series. Sample dilutions are made in 1X Cell Extraction Buffer PTR.
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Sandwich ELISA - Human Fibronectin ELISA Kit (ab219046)To learn more about the advantages of recombinant antibodies see here.
