Human CCL25/TECK DuoSet ELISA Summary

Product Summary

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human CCL25/TECK. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Preparation and Storage

Background: CCL25/TECK

CCL25/TECK is a CC chemokine is produced by stromal cells in the thymus and epithelial cells of the small intestine, particularly the jejunum and ileum. It binds to and induces chemoattraction through CCR9 on immature pre-T cells and thymocytes. CCL25 induces the homing of several lymphocyte populations to the small intestine, including Integrin a4b7+ gamma/delta T cells, Integrin aEb7+ CD8+ T cells, and IgA-producing plasma cells. In cancer, CCL25 promotes the metastasis of melanoma cells to the small intestine, the migration and invasion of some breast carcinomas, and the attraction of mesenchymal stromal cells to multiple myelomas. CCL25 attracts CCR9+ monocytes and macrophages in rheumatoid arthritis and CCR9+ macrophages in atherosclerosis.

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.