Human Fibronectin Quantikine ELISA Kit

  
  • 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.
Control Available
QC200, Quantikine Immunoassay Control Set 834 for Human Fibronectin - Please Inquire
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
Sample123123
n202020202020
Mean2458.112323.356.5115
Standard Deviation1.42.786.351.312.986.38
CV%5.84.85.25.65.35.5

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 Quantikine ELISA Kit
Preparation and Storage
  • 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
      Fibronectin; FN1;
    Related Research Areas
    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.
    ELISA Controls
    Description Application Cat# Citations Images  

    Quantikine Immunoassay Control Set 834 for Human Fibronectin

    Ctrl QC200
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    Supplemental ELISA Products
    Description Application Cat# Citations Images  

    Quantikine ELISA Wash Buffer 1

    ELISA WA126 6
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