Human CD200 DuoSet ELISA Summary

Product Summary

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human CD200. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Preparation and Storage

Background: CD200

CD200 antigen, also known as OX-2, is a type I transmembrane protein belonging to the Ig superfamily. It has a 202 amino acid extracellular domain and a short (19 aa) cytoplasmic tail. At least two alternate splice isoforms exist that differ in their cytoplasmic domains. CD200 is widely expressed in multiple cell types. It interacts with CD200 R1, another Ig superfamily inhibitory receptor primarily expressed on leukocytes of the myeloid lineage, to inhibit myeloid functions.

CD200 R1, also known as OX-2 receptor, is a transmembrane glycoprotein and member of the immunoglobulin superfamily. It is expressed primarily in cells of the myeloid lineage, while its ligand, CD200, is widely distributed. CD200 R1 is though to propagate inhibitory signals and disruption of the CD200/CD200 R1 receptor-ligand pair leads to a predisposition to autoimmune phenotypes.

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.