Human IL-13 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human IL-13 Immunoassay is a 4.5 hour solid phase ELISA designed to measure IL-13 levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human IL-13 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate recombinant human IL-13 accurately. Results obtained using natural human IL-13 showed dose response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural human IL-13.

Preparation and Storage

Background: IL-13

IL-13 (Interleukin-13) plays a key role in the pathogenesis of allergy, cancer, and tissue fibrosis. It is secreted by several helper T cell subsets, NK cells, mast cells, eosinophils, basophils, and visceral smooth muscle cells. IL-13 suppresses the production of proinflammatory cytokines and other cytotoxic substances by macrophages, fibroblasts, and endothelial cells. It promotes B cell activation, immunoglobulin class switching to IgE, and the upregulation of CD23/Fc epsilon RII. IL-13 signals through a receptor complex containing IL-13 R alpha 1 and IL-4 R alpha. This complex also functions as the type 2 IL-4 receptor. IL-13 R alpha 2 binds IL-13 and prevents IL-13 from signaling through the IL-13 R alpha 1/IL-4 R alpha complex. IL-13-bound IL-13 R alpha 2 can directly promote tumor cell invasiveness and the development of tissue fibrosis.

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.