Human CCL11/Eotaxin Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human Eotaxin Immunoassay is a 3.5 hour solid phase ELISA designed to measure human Eotaxin in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human Eotaxin and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate recombinant human Eotaxin accurately. Results obtained measuring natural human eotaxin showed dose-response 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 Eotaxin.

Preparation and Storage

Background: CCL11/Eotaxin

CCL11/Eotaxin is a chemokine that interacts with CCR3 to induce eosinophil, basophil, dendritic cell, and Th2 cell chemoattraction during inflammatory and allergic reactions. Its production is upregulated in airway epithelium, alveolar macrophages, eosinophils, smooth muscle cells, chondrocytes, and fibroblasts following allergen or inflammatory challenge. CCL11 is also involved in the growth of myeloid cell progenitors and the differentiation of mast cells.

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.