Human CCL22/MDC Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human MDC Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human MDC in cell culture supernates, serum, or plasma. It contains E. coli-expressed recombinant human MDC and has been shown to accurately quantitate the recombinant factor. Results obtained using naturally expressed human MDC 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 expressed human MDC.

Preparation and Storage

Background: CCL22/MDC

Macrophage-derived chemokine (MDC), also named stimulated T cell chemotactic protein (STCP-1) and ABCD-1, and now designated as CCL22, is a CC chemokine initially isolated from clones of monocyte-derived macrophages. At the amino acid sequence level, MDC shows less than 35% identity to other CC chemokine family members. Human MDC is expressed in dendritic cells, macrophages and activated monocytes. In addition, MDC expression is detected in thymus, lymph node and appendix tissues. At the amino acid sequence level, mouse and human MDC share 64% identity and 83% similarity.

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.