Human IL-18 BPa Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human IL-18 BPa Immunoassay is a 4.5 hour solid phase ELISA designed to measure IL-18 BPa in cell culture supernates, cell lysates, serum, and plasma. It contains NS0-expressed recombinant human IL-18 BPa and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate recombinant human IL-18 BPa. Results obtained using natural human IL-18 BPa showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human IL-18 BPa.

Preparation and Storage

Background: IL-18 BPa

Interleukin 18 binding proteins are secreted glycoproteins which antagonize IL-18 activity by binding IL-18 and blocking its biological activity. Alternative splicing of the IL-18 BP gene produces at least four isoforms that differ in affinity for IL-18. The IL-18 BP isoforms a and c each contain one immunoglobulin (Ig)-like C2-type domain while isoforms b and d lack a complete Ig-like domain.

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.