Human Fibronectin Quantikine ELISA Kit Summary

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.

Preparation and Storage

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.

Assay Procedure

Refer to the product datasheet for the 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.     Add 50 μL of Assay Diluent to each well.

4.     Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.

5.     Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

6.     Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

7.     Aspirate and wash 5 times.

8.     Add 100 μL Substrate Solution to each well.

9.     Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.