Human CD163 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human CD163 immunoassay is a 4.5 hour solid phase ELISA designed to measure human soluble CD163 in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant human CD163 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human CD163 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 CD163.

Preparation and Storage

Background: CD163

CD163, also known as M130 and p155, is a transmembrane scavenger receptor that is expressed on monocytes and macrophages and is inducible by immunosuppressant glucocorticoids and IL-10. A soluble form is shed from the cell surface by TACE or neutrophil elastase mediated cleavage in response to oxidative stress, Prostaglandin F2a stimulation, or the activation of Fc gamma receptors, TLR1, 2, 5, or 6. CD163 mediates monocyte binding to bacteria, leading to the release of inflammatory cytokines. It is essential for the circulatory clearance of hemoglobin-haptoglobin (Hb-Hp) complexes as well as free hemoglobin. It can also mediate monocyte-erythroblast adhesion and promote erythroblast expansion. CD163 binds and internalizes the cytokine TWEAK, and the ratio of soluble CD163 to TWEAK in the plasma is elevated during atherosclerosis.

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.