Human TFF3 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human TFF3 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human TFF3 in cell culture supernates, serum, plasma, saliva, and urine. It contains CHO cell-expressed recombinant human TFF3 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human TFF3 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human TFF3.

Preparation and Storage

Background: TFF3

TFF3 (Trefoil Factor 3), also known as ITF and P1.B, is a peptide that plays an important role in epithelial regeneration and wound healing. It can form disulfide-linked dimers or associate into disulfide-linked complexes with the intestinal mucous proteins FCGBP and MUC-2. TFF3 is expressed by epithelial goblet cells in the respiratory tract, biliary and breast ducts, small and large intestine, and cardia of the stomach as well as by chondrocytes during bone development. TFF3 is upregulated in response to gastrointestinal epithelial disruptions and promotes epithelial wound healing. It enhances tumor cell invasion and metastasis by supporting hypoxia-induced VEGF upregulation.

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.