Human EMMPRIN/CD147 Quantikine ELISA Kit Summary

Product Summary

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

Preparation and Storage

Background: EMMPRIN/CD147

The name EMMPRIN reflects this proteins extracellular metalloproteinase inducer activity. It is a cell surface protein with two extracellular Ig domains, a transmembrane domain, and a cytoplasmic domain. Its expression is often elevated in human tumor cells, which in turn promotes MMP-1, MMP-2 and MMP-3 production in melanoma cells and MMP-2 and MMP-9 production in breast cancer cells. It interacts with several types of molecules such as integrins, caveolin-1 and MCT1. TRA-1-85 is an epitope within EMMPRIN.

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.