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|Method & Line||Sample & Target||Product||Package Info|
|MicroELISA||Serum||EIAgen Free PSA Kit||Tests per Package: 96|
|EIAgen||EIAgen Free PSA Kit is a direct solid phase enzyme immunoassay for the quantitative measurement of Free Prostate Specific Antigen (fPSA) in human serum.||Code: LI4037F1||Package: 1 Microplate|
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EIAgen Free PSA Kit is a direct solid phase enzyme immunoassay for the quantitative measurement of Free Prostate Specific Antigen (fPSA) in human serum.
Prostate Specific antigen (PSA) is a serine protease with chymotrypsin-like activity (1,2). The protein is a single chain glycoprotein with a molecular weight of 28.4 kDA (3). PSA derives its name from the observation that it is a normal antigen of the prostrate but is not found in any other normal or malignant tissue. PSAis released from the normal prostate and appears atlow serum concentrations in healthy men. Studies with reverse transcription-PCRhave shown that PSAalso is expressed at a low concentrationin peripheral blood cells and other tissues (4). High serumconcentrations can be detected in patients with advanced prostatecancer (PCA) (5). Therefore PSA is applied as a tumor markerfor the clinical management of PCA (6). However, increased PSAconcentrations in serum also occur in patients with benign prostatehyperplasia (BPH) (7). Hence the goal is to discriminate clearlybetween BPH and PCA in the clinical laboratory to spare thepatient invasive diagnostic procedures, such as a prostate biopsy.
In human serum PSAoccurs in two forms: free PSA (f-PSA) andcomplexed PSA. The major form is a complex of PSA and α1-antichymotrypsin(ACT). The fraction of f-PSA was shown to be substantially smallerin patients with untreated PCA than in patients with BPH. Thereforecombined measurements of f-PSAand total PSA (t-PSA) may leadto a better discrimination between BPH and PCA Somerecent studies have already shown that the f-PSA/t-PSA ratiois helpful in the differential diagnosis of BPH and PCA.
PSA is found in benign, malignant and metastatic prostrate cancer. Since prostate cancer is the second most prevalent form of male malignancy, the detection of elevated PSA levels plays an important role in the early diagnosis. Serum PSA levels have been found to be more useful than prostatic acid phosphatase (PAP) in the diagnosis and management of patients due to increased sensitivity (4).
In this method, fPSA calibrator, patient specimen or control is first added to a streptavidin coated well. Biotinylated monoclonal and enzyme labeled antibodies (directed against distinct and different free epitopes of fPSA) are added and the reactants mixed. Reaction between the various PSA antibodies and native PSA forms a sandwich complex that binds with the streptavidin coated to the well.
After the completion of the required incubation period, the enzyme-fPSA antibody bound conjugate is separated from the unbound enzyme-fPSA conjugate by aspiration or decantation. The activity of the enzyme present on the surface of the well is quantitated by reaction with a suitable substrate to produce color.
The employment of several serum references of known prostate specific antigen (fPSA) levels permits the construction of a dose response curve of activity and concentration. From comparison to the dose response curve, an unknown specimen's activity can be correlated with fPSA concentration.
The essential reagents required for an immunoenzymometric assay include high affinity and specificity antibodies (enzyme and immobilized), with different and distinct epitope recognition, in excess, and native antigen. In this procedure, the immobilization takes place during the assay at the surface of a microplate well through the interaction of streptavidin coated on the well and exogenously added biotinylated monoclonal anti-PSA antibody.
Upon mixing monoclonal biotinylated antibody, the enzyme-labeled antibody and a serum containing the native antigen, reaction results between the native antigen and the antibodies, without competition or steric hindrance, to form a soluble sandwich complex.
Simultaneously, the complex is deposited to the well through the high affinity reaction of streptavidin and biotinylated antibody.
After equilibrium is attained, the antibody‑bound fraction is separated from unbound antigen by decantation or aspiration. The enzyme activity in the antibody‑bound fraction is directly proportional to the native antigen concentration. By utilizing several different serum references of known antigen values, a dose response curve can be generated from which the antigen concentration of an unknown can be ascertained.
The kit contains reagents for 96 tests (code LI4037F1).
Bring reagents to room temperature before use.
Wash Buffer Conc. 50X
Number of tests