Human Fibronectin ELISA Kit (ab219046)

Human Fibronectin ELISA Kit is a single-wash 90-min Simplestep used to quantify Human Fibrionectin with a sensitivity of 20.6 pg/mL. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Different formats for different needs: 384-well plate for higher throughput
- Design your own immunoassay: we also offer the conjugation-ready antibody pair

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Key facts

Detection method: Colorimetric
Sample types: Cell culture extracts, Tissue Extracts, Citrate plasma, Cell culture supernatant, Milk, Serum, EDTA Plasma
Assay type: Sandwich (quantitative)
Reactive species: Human, Rhesus monkey
Range: 125 - 8000 pg/mL
Assay time: 1h 30m
Sensitivity: = 20.6 pg/mL

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Reactivity data

Application	Reactivity	Dilution info	Notes
Application sELISA	Reactivity Reacts	Dilution info -	Notes -

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Target data

See full target information FN1

Function

Fibronectins bind cell surfaces and various compounds including collagen, fibrin, heparin, DNA, and actin (PubMed:3024962, PubMed:3593230, PubMed:3900070, PubMed:7989369). Fibronectins are involved in cell adhesion, cell motility, opsonization, wound healing, and maintenance of cell shape (PubMed:3024962, PubMed:3593230, PubMed:3900070, PubMed:7989369). Involved in osteoblast compaction through the fibronectin fibrillogenesis cell-mediated matrix assembly process, essential for osteoblast mineralization (By similarity). Participates in the regulation of type I collagen deposition by osteoblasts (By similarity). Acts as a ligand for the LILRB4 receptor, inhibiting FCGR1A/CD64-mediated monocyte activation (PubMed:34089617). 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. Secreted by contracting muscle, induces liver autophagy, a degradative pathway for nutrient mobilization and damage removal, and systemic insulin sensitization via hepatic ITGA5:ITGB1 integrin receptor signaling.

Alternative names

FN, FN1, Fibronectin, Cold-insoluble globulin, CIG

What's included?

1 x 384 Tests

Components

10X Human Fibronectin Capture Antibody

1 x 600 µL

10X Human Fibronectin Detector Antibody

1 x 600 µL

10X Wash Buffer PT (ab206977)

2 x 20 mL

384 well CaptSure™ microplates

1 x 1 Unit

50X Cell Extraction Enhancer Solution (ab193971)

6 x 1 mL

5X Cell Extraction Buffer PTR (ab193970)

1 x 50 mL

Antibody Diluent 4BR

1 x 6 mL

Human Fibronectin Lyophilized Recombinant Protein

2 x 1 Vial

Plate Seals

1 x 1 Unit

Sample Diluent NS (ab193972)

2 x 50 mL

Stop Solution

2 x 12 mL

TMB Development Solution

2 x 12 mL

Key facts

Detection method: Colorimetric
Sample types: Cell culture extracts, Tissue Extracts, Citrate plasma, Cell culture supernatant, Milk, Serum, EDTA Plasma
Assay type: Sandwich (quantitative)
Reactive species: Human, Rhesus monkey
Range: 125 - 8000 pg/mL
Assay time: 1h 30m
Assay Platform: Pre-coated microplate (12 x 8 well strips)
Sensitivity: = 20.6 pg/mL

Precision

Intra assay

Sample	n	C.V.
Sample Serum	n 3	C.V. 5

Inter assay

Sample	n	C.V.
Sample Serum	n 5	C.V. 8.3

Recovery

Sample specific recovery

Sample type	Average %	Range
Sample type Cell culture supernatant	Average % = 102	Range 90 - 117 %
Sample type Milk	Average % = 102	Range 91 - 111 %
Sample type Serum	Average % = 93	Range 87 - 102 %
Sample type EDTA Plasma	Average % = 86	Range 81 - 94 %
Sample type Cell culture extracts	Average % = 89	Range 80 - 105 %
Sample type Tissue Extracts	Average % = 90	Range 80 - 131 %
Sample type Citrate plasma	Average % = 99	Range 87 - 120 %

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C
Storage information: +4°C

Notes

Human Fibronectin ELISA Kit ab219046 is a rapid single-wash 90-min ELISA kit to measure Human Fibronectin in cell culture extracts, tissue extracts, cell culture supernatant, milk, serum and plasma. This SimpleStep ELISA® sensitivity is 20.6 pg/mL.

How the assay works

Human Fibronectin SimpleStep ELISA® 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.

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.
Recombinant human Anastelin was prepared within the working range of the assay and assayed for cross reactivity. No cross-reactivity was observed.

Human Fibronectin ELISA Kit ab219046 protocol summary:

1. Mix: add samples/standards to the wells together with the capture and detector antibody cocktail. Incubate for 1 hr at room temperature
2. Wash
3. Add TMB development solution - incubate for 10 min
4. Add stop solution
5. Read the results on a plate reader at 450nm

How other researchers are using Human Fibronectin ELISA Kit ab219046

Human Fibronectin ELISA Kit ab219046 has been used to study wound healing after dermato-oncological surgery ⁽¹⁾ and the association of NGAL and fibrosis in diabetic nephropathy⁽²⁾.

References:

⁽¹⁾Cristina Magnoni et al. 2023, PMID: 36836820,
⁽²⁾Guiying Li et al. 2021, PMID: 33732294

Save your precious samples

Our 384-well Human Fibronectin ELISA Kit only requires a maxmum of 12.5µL

Design your own immunoassay

We offer the antibody pair used in this kit in a BSA and Azide-free format, ready for conjugation

- Human Fibronectin Antibody Pair - BSA and Azide free Human Fibronectin Antibody Pair - BSA and Azide free ab241837
- Anti-Fibronectin antibody [EPR19331-123] - BSA and Azide free (Capture) Anti-Fibronectin antibody [EPR19331-123] - BSA and Azide free (Capture) ab242680
- Anti-Fibronectin antibody [EPR19331-44] - BSA and Azide free (Detector) Anti-Fibronectin antibody [EPR19331-44] - BSA and Azide free (Detector) ab242925

Related and recommended products

Human Fibronectin ELISA Kit ab219046 is commonly used to study tissue structure and function. Fibronectin is a component of the extracellular matrix (ECM).

Other ELISA kits to measure the ECM and its role in tissue function include:

- Human Pro-Collagen I alpha 1 ELISA kit Human Pro-Collagen I alpha 1 ELISA Kit ab210966
- Human alpha SMA ELISA Kit Human alpha SMA ELISA Kit ab240678
- Human MCP-1 ELISA Kit Human MCP-1 ELISA Kit ab179886
- Human Elastin ELISA kit Human Elastin ELISA Kit ab239433

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.

REACH authorisation
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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.

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20 product images

ELISA - Human Fibronectin ELISA Kit (ab219046)
Sandwich ELISA - Human Fibronectin ELISA Kit (ab219046)
Example of human Fibronectin standard curve in Sample Diluent NS.
Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Human Fibronectin ELISA Kit (ab219046)
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).
Sandwich ELISA - Human Fibronectin ELISA Kit (ab219046)
Example of human Fibronectin standard curve in 1X Cell Extraction Buffer PTR.
Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Human Fibronectin ELISA Kit (ab219046)
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.
ELISA - Human Fibronectin ELISA Kit (ab219046)
Human Fibronectin standard curve comparison data.
Standard curve comparison between human Fibronectin SimpleStep ELISA^® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows comparable sensitivity.
Sandwich ELISA - Human Fibronectin ELISA Kit (ab219046)
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.
Sandwich ELISA - Human Fibronectin ELISA Kit (ab219046)
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.
Sandwich ELISA - Human Fibronectin ELISA Kit (ab219046)
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.
Sandwich ELISA - Human Fibronectin ELISA Kit (ab219046)
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.
Sandwich ELISA - Human Fibronectin ELISA Kit (ab219046)
Assay sensitivity.
The MDD was determined by calculating the mean of zero standard replicates and adding 2 standard deviations then extrapolating the corresponding concentration.
Sandwich ELISA - Human Fibronectin ELISA Kit (ab219046)
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.
Sandwich ELISA - Human Fibronectin ELISA Kit (ab219046)
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.
Sandwich ELISA - Human Fibronectin ELISA Kit (ab219046)
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.
Sandwich ELISA - Human Fibronectin ELISA Kit (ab219046)
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.
Sandwich ELISA - Human Fibronectin ELISA Kit (ab219046)
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.
Sandwich ELISA - Human Fibronectin ELISA Kit (ab219046)
Example of human Fibronectin standard curve in Sample Diluent NS in 96-well vs. 384-well plate.
Example of human Fibronectin standard curve in 96-well vs. 384-well plate. Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Human Fibronectin ELISA Kit (ab219046)
Interpolated concentrations of human Fibronectin in HepG2 cell extract in 96-well vs. 384-well plates.
Interpolated concentration of native Fibronectin was measured in duplicate at different sample concentrations in 96-well vs. 384-well plates. Undiluted samples are 25 µg/mL HepG2 cell extract. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in 1X Cell Extraction Buffer PTR.
Sandwich ELISA - Human Fibronectin ELISA Kit (ab219046)
Interpolated concentrations of human Fibronectin in serum in 96-well vs. 384-well plates.
Interpolated concentration of native Fibronectin was measured in duplicate at different sample concentrations in 96-well vs. 384-well plates. Undiluted samples are 1:20,000 serum. The interpolated dilution factor corrected values (to neat sample) are plotted (mean +/- SD, n=2). Sample dilutions are made in Sample Diluent NS.
Sandwich ELISA - Human Fibronectin ELISA Kit (ab219046)
Example of human Fibronectin standard curve in 1X Cell Extraction Buffer PTR in 96-well vs. 384-well plate.
Example of human Fibronectin standard curve in 96-well vs. 384-well plate. Background-subtracted data values (mean +/- SD) are graphed.