Human CD31/PECAM-1 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human CD31/PECAM-1 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human CD31 in cell culture supernates, serum, plasma, saliva, and urine. It contains CHO cell-expressed recombinant human CD31 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human CD31 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 CD31.

Preparation and Storage

Background: CD31/PECAM-1

CD31/PECAM-1 is a heavily glycosylated transmembrane homophilic adhesion protein that is highly expressed on endothelial cells and at a lower level on platelets, granulocytes, macrophages, dendritic cells, T and B cells, and NK cells. It is required for the transepithelial migration of leukocytes. CD31 signaling increases the activation threshold of T cells and inhibits dendritic cell development. The extracellular domain of CD31 is released during endothelial cell apoptosis. This fragment circulates in the serum of patients suffering from myocardial infarction, acute ischaemic stroke, and multiple sclerosis, conditions that involve tissue damage and endothelial cell apoptosis.

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.