Human ICAM-1/CD54 Allele-specific Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human sICAM-1 immunoassay is a 2.0 hour solid phase ELISA designed to measure sICAM-1 in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant human sICAM-1 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human sICAM-1 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 sICAM-1.

Preparation and Storage

Background: ICAM-1/CD54

ICAM-1 (intercellular adhesion molecule-1), also known as CD54, is a transmembrane protein that is upregulated on endothelial and epithelial cells at sites of inflammation. It mediates the vascular adhesion and paracellular migration of leukocytes expressing activated LFA-1 (CD11a/CD18) and Mac-1 (CD11b/CD18). It also binds several non-integrin ligands including CD43/Sialophorin, Fibrinogen, Hyaluronan, rhinoviruses, and Plasmodium falciparum-infected erythrocytes. Soluble ICAM-1 promotes angiogenesis and serves an indicator of vascular endothelial cell activation or damage. Elevated levels of soluble ICAM-1 are associated with cardiovascular disease, type 2 diabetes, organ transplant dysfunction, oxidant stress, increased abdominal fat mass, hypertension, liver disease, certain malignancies, and cerebral malaria.

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.