Human CCL17/TARC Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human TARC Immunoassay kit is a 3.5 hour solid phase ELISA designed to measure TARC in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human TARC and antibodies raised against the recombinant factor. It has been shown to accurately quantitate recombinant human TARC. Results obtained using natural human TARC showed linear curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human TARC.

Preparation and Storage

Background: CCL17/TARC

CCL17/TARC is a chemokine that interacts with CCR4 and CCR8 to induce the chemoattraction of activated Th2 cells, basophils, and NK cells. It is produced by thymic dendritic cells, lymph node dendritic cells, monocytes, CD4+ T cells, keratinocytes, fibroblasts, bronchial epithelial cells, and Reed-Sternberg cells. CCL17 promotes Th2 cell recruitment to sites of inflammation and induces platelet aggregation and degranulation.

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.