Human Fibronectin Quantikine ELISA Kit

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DFBN10
Control Products Available
Human Fibronectin ELISA Standard Curve
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Product Details
Procedure
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Human Fibronectin Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Cell Lysates (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Citrate Plasma (10 uL), Saliva (17 uL), Urine (17 uL), Human Milk (10 uL)
Sensitivity
0.579 ng/mL
Assay Range
3.1 - 200 ng/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma, Saliva, Urine, Human Milk)
Specificity
natural and recombinant human Fibronectin and Fibronectin fragments aa 1-1908, aa 607-1265, aa 627-702. Fibronectin fragment aa 631-705 cross-reacts approximately 0.415% Fibronectin fragments aa 1266-1908, aa 1722-1811, and aa 1913-2477 are not detected in this kit.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Human Fibronectin Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Fibronectin in serum-free cell culture supernates, cell lysates, tissue lysates, serum, plasma, saliva, urine, and human milk.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components

Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma, Saliva, Urine, Human Milk

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 24 58.1 123 23.3 56.5 115
Standard Deviation 1.4 2.78 6.35 1.31 2.98 6.38
CV% 5.8 4.8 5.2 5.6 5.3 5.5

Recovery

The recovery of human Fibronectin spiked to levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 97 89-115

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human Fibronectin were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay. Samples were diluted prior to assay.
Human Fibronectin ELISA Linearity

Data Examples

Human Fibronectin ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Fibronectin

Fibronectin is an extracellular matrix (ECM) component and is one of the primary cell adhesion molecules. The occurrence of different isoforms is due to alternative mRNA splicing of the ED-A, ED-B and III-CS regions, and subsequent post-translational modification. Although non-reactive with adhesion receptors in its soluble state, fibronectin is highly adhesive when immobilized on a surface.

Entrez Gene IDs:
2335 (Human); 280794 (Bovine)
Alternate Names:
CIG; ED-B; fibronectin 1; Fibronectin; FINC; FN; FN1; FNZ; GFND; GFND2; LETS; MSF; SMDCF
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

FAQs

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Human Fibronectin Quantikine ELISA Kit
By Anonymous on 01/31/2019
Application: Sample Tested: Serum and Plasma