Mouse IL-23 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Mouse IL-23 immunoassay is a 4.5 hour solid-phase ELISA designed to measure IL-23 in cell culture supernates, mouse serum, and plasma. It contains Sf 21-expressed recombinant mouse IL-23 and antibodies raised against the recombinant p19 and p40 subunits. This immunoassay has been shown to accurately quantitate the recombinant mouse IL-23. Results obtained using natural mouse IL-23 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 IL-23.

Preparation and Storage

Background: IL-23

IL-23 (Interleukin-23) is a disulfide-linked cytokine composed of a p19 subunit that is unique to IL-23 and a p40 subunit that is shared with IL-12. It is produced by activated macrophages, microglia, and monocyte-derived dendritic cells in response to pathogens. IL-23 induces the earliest recruitment of neutrophils to the site of infection and promotes the development and maintenance of Th17 cells. It also enhances the development of Th17 mediated autoimmunity and tumor progression. IL-23 signals through a receptor complex consisting of IL-12 R beta 1 and IL-23 R. This complex is expressed in mouse Th1 and Th2 cells, bone marrow dendritic cells, activated macrophages and CD4+ CD45Rb(low) memory T 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.