Human CXCL16 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human CXCL16 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human CXCL16 in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human CXCL16 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human CXCL16 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 naturally occurring human CXCL16.

Preparation and Storage

Background: CXCL16

CXC chemokine ligand 16 (CXCL16) is a type I membrane protein that has a non-ELR CXC chemokine domain in its extracellular region. Fractalkine (CX3CL1) and CXCL16 constitute the only two transmembrane chemokines. Mouse and human CXCL16 share 70% amino acid sequence similarity within their chemokine domains and 49% overall amino acid sequence identity.

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.