Human COMP Quantikine ELISA Kit Summary

Product Summary

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

Preparation and Storage

Background: COMP/Thrombospondin-5

COMP (Cartilage Oligomeric Matrix Protein), also known as Thrombospondin-5, is a multidomain calcium binding protein that associates with other extracellular matrix molecules. COMP associates into disulfide-linked homopentamers with a central hub and peripheral globular domains connected by flexible strands. An axial pore is formed by the coiled coil assembly and binds vitamin D3 which is involved in bone and cartilage metabolism. An RGD sequence in the third TSP type-3 repeat mediates chondrocyte attachment via Integrins a5b1 or aVb3. COMP is upregulated in rheumatoid arthritis and osteoarthritis, hepatocellular carcinomas, chronic pancreatitis, and pancreatic carcinomas. Several mutations in COMP are associated with skeletal dysplasias.

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.