Human PSA ELISA Kit

RACTIVITY	Human
SENSITIVITY	<1.0 ng/mL
ASSAY RANGE	1-80 ng/mL
REAGENTS PROVIDED	PSA MICROTITER PLATE  PSA CONJUGATE  PSA STANDARD - 80 ng/mL PSA STANDARD - 40 ng/mL PSA STANDARD - 20 ng/mL PSA STANDARD - 10 ng/mL PSA STANDARD - 2 ng/mL PSA STANDARD - 0 ng/mL SUBSTRATE A  SUBSTRATE B  STOP SOLUTION SAMPLE DILUENT

INTENDED USE

This Human PSA Kit ELISA Kit is to be used for the in vitro quantitative determination of human PSA concentrations in serum and cell culture samples.  This kit is intended for LABORATORY RESEARCH USE ONLY and is not for use in diagnostic or therapeutic procedures.

INTRODUCTION

Commonly known as Prostate-Specific Antigen (PSA), Kallikrein-3 is a peptidase that is produced in large amount by epithelial cells of the male prostate gland and female paraurethral duct during ejaculation.  The protein can be produced by normal, benign, and cancerous cells.  In circulation, majority of kallikrein-3 forms complex with a1-anti-chymotrypsin, with minor amount binding to other proteins.   Kallikrein-3 serum levels are elevated in patients with prostate cancer, benign prostatic hypertrophy (BPH) and inflammatory conditions associated with the tissue.  Kallikrein-3 has been chosen as serum marker for prostate cancer screening for males over 50 years older.  Recently, this application has been challenged because its potential benefit may be out-weighted by over-diagnosis and overtreatment.  In molecular biological research, it was found that the expression of Kallikrein-3 is upregulated by androgen receptor signalling pathway and down-regulated by EGF signalling pathway.  The integrity of androgen receptor pathway can be monitored by measuring kallikrein-3 concentrations in biological concentrations and cell supernatant. 

PRINCIPLE OF THE ASSAY

This PSA enzyme linked immunosorbent assay (ELISA) applies a technique called a quantitative sandwich immunoassay.  The microtiter plate provided in this kit has been pre-coated with a monoclonal antibody specific for PSA.  Standards or samples are then added to the microtiter plate wells and PSA, if present, will bind to the antibody pre-coated on the wells.  In order to quantify the amount of PSA present in the sample, a standardized preparation of horseradish peroxidase (HRP)-conjugated monoclonal antibody, specific for PSA are added to each well to “sandwich” the PSA immobilized on the plate. The microtiter plate then undergoes incubation, followed by thorough washing of the wells to remove all unbound components.  Next, a TMB (3,3',5,5' tetramethyl-benzidine) substrate solution is added to each well.  The enzyme (HRP) and substrate are allowed to react over a short incubation period.  Only those wells that contain PSA and enzyme-conjugated antibody will exhibit a change in colour.  The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the colour change is measured spectrophotometrically at a wavelength of 450nm ± 2nm.

In order to measure the concentration of PSA in the sample, this Human PSA ELISA Kit includes a set of calibration standards (6 standards).  The calibration standards are assayed at the same time as the samples allowing the operator to produce a standard curve of Optical Density (O.D.) versus PSA concentration (ng/mL).  The concentration of PSA in the samples is then determined by comparing the O.D. of the samples to the standard curve.

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