Human IL-11 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human IL-11 Immunoassay is a 4.5-5.0 hour, solid-phase ELISA designed to measure IL-11 in cell culture supernates, serum, or plasma. It contains Sf 21-expressed recombinant human IL-11 and antibodies raised against recombinant human IL-11. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural human IL-11 showed dose 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 IL-11.

Preparation and Storage

Background:  IL-11

Interleukin-11, also known as AGIF, is a pleiotropic cytokine in the IL-6 family, which also includes LIF, CNTF, Oncostatin M, Cardiotrophin-1, IL-27, and IL-31. It is secreted by osteoblasts, synoviocytes, fibroblasts, chondrocytes, intestinal myofibroblasts, and trophoblasts. It is found in the plasma mainly during inflammation, and is considered to be primarily anti-inflammatory. It stimulates hematopoiesis and thrombopoiesis, regulates macrophage differentiation, and confers mucosal protection in the intestine. It has also been found to enhance T cell polarization toward Th2, promote B cell IgG production, increase osteoclast bone absorption, protect endothelial cells from oxidative stress, and regulate epithelial proliferation and apoptosis. IL-11 signals through a receptor complex that contains IL-11 R alpha and gp130. A soluble form of IL-11 R alpha can bind IL-11 and either form a signaling complex with gp130 on the cell surface or inhibit cell surface IL-11 R alpha/gp130 signaling.

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.