Human CA125/MUC16 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human CA125/MUC16 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human CA125 in cell culture supernates, serum, plasma, saliva, urine, and human milk. It contains CHO cell-expressed recombinant human CA125 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human CA125 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 CA125.

Preparation and Storage

Background: CA125/MUC16

The CA125 antigen, also known as MUC16, is a mucin protein that may be found in type I transmembrane or secreted forms that are used monitor the progress of epithelial ovarian cancer therapy. Expression of isoforms, proteolytic cleavage, and heavy N- and O- linked glycosylation produce forms of human CA125 that can vary from 200-5000 kDa in size. It inhibits NK cell activity, which is thought to contribute to immune evasion in peritoneal cancer and pregnancy.

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.