Human Mesothelin Quantikine ELISA Kit Summary

Product Summary

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

Preparation and Storage

Background: Mesothelin

Mesothelin, also known as CAK1 and ERC, is derived from a precursor that also includes Megakaryocyte Potentiating Factor (MPF). Following cleavage of the precursor, Mesothelin remains attached to the cell surface via a GPI linkage. Mesothelin is normally expressed on mesothelial cells in the pleura, pericardium, and peritoneum as well as in the developing and postnatal pancreas. It is upregulated in mesotheliomas and a range of carcinomas and adenomas. Mesothelin promotes tumor cell proliferation, migration, anchorage-independent growth, and tumor progression.

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.