Mouse CCL21/6Ckine Quantikine ELISA Kit Summary

Product Summary

The Quantikine Mouse CCL21/6Ckine Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse 6Ckine in tissue culture supernates, serum, and plasma. It contains E. coli-expressed recombinant mouse 6Ckine and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant mouse 6Ckine. Results obtained using natural mouse 6Ckine showed dose response curves that were parallel to the standard curves obtained using the Quantikine mouse kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse 6Ckine.

Preparation and Storage

Background: CCL21/6Ckine

CCL21, also known as 6Ckine, TCA-4, SLC, Exodus-2, and A21, is a homeostatic chemokine that signals through CCR7 or CXCR3. CCL21 is constitutively presented on initial lymphatic vessels, high endothelial venules (HEV), and lymph node dendritic cells (DC) where it promotes the docking of DC to lymphatic vessels and the retention of T cells by lymph node DC, resulting in T cell priming for activation. Its upregulation during chronic inflammation or tissue damage promotes fibrosis, inflammatory cytokine production, and neuropathic pain. The soluble chemokine is elevated in rheumatoid arthritis synovial fluid and in the serum of coronary artery disease patients.

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.