Mouse LIX Quantikine ELISA Kit Summary

Product Summary

The Quantikine Mouse CXCL5/LIX Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse LIX levels in cell culture supernates, tissue lysates, serum, platelet-poor plasma, and urine. It contains E. coli-expressed recombinant mouse LIX and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant mouse LIX accurately. Results obtained using natural mouse LIX 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 mouse LIX.

Preparation and Storage

Background: LIX

LPS-induced CXC chemokine (LIX) is a member of the murine chemokine family. It is 132 amino acids (aa) in length with a predicted molecular weight of 14 kDa prior to cleavage of the N-terminal signal peptide (aa 1–40). LIX has no true human ortholog but is closely related to human CXCL5/ENA-78 and CXCL6/GCP-2. It is widely expressed, and is secreted by multiple cell types including fibroblasts, thymic epithelium, platelets, vascular endothelium, and hepatocytes. LIX acts as a proinflammatory chemokine and is chemotactic for neutrophils.

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.