Human Kallikrein 3/PSA Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human KLK3/PSA Immunoassay is a 4.5 hour solid phase immunoassay designed to measure KLK3/PSA in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant human KLK3/PSA, and antibodies raised against the recombinant protein. Natural human KLK3/PSA showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards, indicating that this kit can be used to determine relative levels of natural human KLK3/PSA.

Preparation and Storage

Background: Kallikrein 3/PSA

The human tissue kallikrein (KLK) gene family contains 15 members that play important roles in cancer. Notably, kallikrein-1, also known as tissue kallikrein, cleaves kininogen to release the vasoactive kinin peptide, bradykinin or lysyl-bradykinin. Kallikrein-3, called prostate specific antigen (PSA), is an established tumor marker that aids in the diagnosis, staging, and follow up of prostate cancer. Kallikrein-4 is specifically expressed in the prostate and over-expressed in prostate cancer. Kallikrein-5 is widely expressed but found at high levels in skin, breast, brain and testis; over-expression is an indicator of poor prognosis in ovarian cancer. Kallikrein-8 is expressed in the brain and is a novel marker of ovarian and cervical cancer.

Human plasma kallikrein, a serine protease, is synthesized in the liver and circulates in the plasma bound to high molecular weight (HMW) kininogen or as a free zymogen. Once activated by its physiological activator, coagulation factor XII, it displays endopeptidase activity towards peptide bonds after arginine (preferred) and lysine. It cleaves HMW kininogen, its major physiological substrate, to release the potent vasodilator peptide bradykinin. It is also able to cleave a number of inactive precursor proteins to generate active products, such as plasminogen and prourokinase

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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.