Mouse IL-15R alpha DuoSet ELISA Summary

Product Summary

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant mouse IL-15 R alpha. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Preparation and Storage

Background: IL-15R alpha

IL-15 Ra (Interleukin 15 Receptor alpha), also known as CD215, is a transmembrane glycoprotein that plays an important role in NK and CD8+ memory T cell homeostasis and activation. It transmits IL-15 induced signals and additionally interacts with a complex of IL-2 R beta and the common gamma chain (gammaC) which are also subunits of the IL-2 receptor complex. In transpresentation, IL-15/IL-15 Ra complexes are expressed on the surface of one cell and interact with complexes of IL-2 R beta/gammaC on adjacent cells. IL-15/IL-15 Ra complexes can transmit reverse signaling that promotes adhesion and activation of the IL-15/IL-15 Ra expressing cells. Shed soluble forms of IL-15 Ra retain the ability to bind tightly to IL-15 and can inhibit IL-15 bioactivity.

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.