Free PSA Kit CLIA

N° du produit ABIN504769

Détail du produit Free PSA Kit CLIA

Antigène: Free PSA (fPSA) (Free Prostate Specific Antigen (fPSA))

Reactivité: Humain

Méthode de détection: Chemiluminescent

Type de méthode: Sandwich ELISA

Application: ELISA

Fonction: Immunoenzymometric assay (TYPE 3): 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.

Analytical Method: Quantitative

Attributs du produit: The Quantitative Determination of Free Prostrate Specific Antigen (fPSA) Concentration in Human Serum by a Microplate Chemiluminescence Immunoassay .

Information d'application

Indications d'application: All products that contain human serum have been found to be non-reactive for Hepatitis B Surface Antigen, HIV 1&2 and HCV Antibodies by FDA required tests. Since no known test can offer complete assurance that infectious agents are absent, all human serum products should be handled as potentially hazardous and capable of transmitting disease. Good laboratory procedures for handling blood products can be found in the Center for Disease Control / National Institute of Health, "Biosafety in Microbiological and Biomedical Laboratories," 2nd Edition, 1988, HHS Publication No. (CDC) 88-8395.

Volume d'échantillon: 25 μL

Plaque: Pre-coated

Protocole:

Specimien Collection and Preparation:

The specimens shall be blood, serum in type and the usual precautions in the collection of venipuncture samples should be observed. For accurate comparison to established normal values, a fasting morning serum sample should be obtained. The blood should be collected in a plain redtop venipuncture tube without additives or anti-coagulants. Allow the blood to clot. Centrifuge the specimen to separate the serum from the cells. Samples may be refrigerated at 2_x001E_8C for a maximum period of five (5) days. If the specimen(s) cannot be assayed within this time, the sample(s) may be stored at temperatures of _x001E_20C for up to 30 days. Avoid repetitive freezing and thawing. When assayed in duplicate, 0.100ml of the specimen is required.

Reagent Preparation:

1. Wash Buffer Dilute contents of Wash Concentrate to 1000ml with distilled or deionized water in a suitable storage container. Store diluted buffer at room temperature 20-27(C. 2. Working Signal Reagent Solution - Store at 2 - 8(C. Determine the amount of reagent needed and prepare by mixing equal portions of Signal Reagent A and Signal Reagent B in a clean container. For example, add 1 ml of A and 1ml of B per two (2) eight well strips (A slight excess of solution is made). Discard the unused portion if not used within 36 hours after mixing. If complete utilization of the reagents is anticipated, within the above time constraint, pour the contents of Signal Reagent B into Signal Reagent A and label accordingly.

Test Procedure:

Before proceeding with the assay, bring all reagents, serum references and controls to room temperature (20 - 27(C). 1. Format the microplates wells for each serum reference, control and patient specimen to be assayed in duplicate. Replace any unused microwell strips back into the aluminum bag, seal and store at 2-8(C. 2. Pipette 0.050 ml (50l) of the appropriate serum reference, control or specimen into the assigned well. 3. Add 0.100 ml (100l) of the fPSA Tracer Reagent to each well. It is very important to dispense all reagents close to the bottom of the coated well. 4. Swirl the microplate gently for 20-30 seconds to mix and cover. 5. Incubate 45 minutes at room temperature. 6. Discard the contents of the microplate by decantation or aspiration. If decanting, tap and blot the plate dry with absorbent paper. 7. Add 350l of wash buffer (see Reagent Preparation Section), decant (tap and blot) or aspirate. Repeat four (4) additional times for a total of five (5) washes. An automatic or manual plate washer can be used. Follow the manufacturers instruction for proper usage. If a squeeze bottle is employed, fill each well by depressing the container (avoiding air bubbles) to dispense the wash. Decant the wash and repeat four (4) additional times. Add 0.100 ml (100l) of working signal reagent (see Reagent Preparation Section) to all wells. Always add reagents in the same order to minimize reaction time differences between wells. Incubate for five (5) minutes at room temperature in the dark. 10. Read the relative light units in each well for 0.2 1.0 seconds. The results should be read within thirty (30) minutes of adding the signal reagent solution.

Restrictions: For Research Use only

Stockage

Détail du antigène

Antigène: Free PSA (fPSA) (Free Prostate Specific Antigen (fPSA))

Autre désignation: Prostate Specific Antigen (PSA) (free) (fPSA Produits)

Sujet: 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. PSA is released from the normal prostate and appears at low serum concentrations in healthy men. Studies with reverse transcription-PCR have shown that PSA also is expressed at a low concentration in peripheral blood cells and other tissues (4). High serum concentrations can be detected in patients with advanced prostate cancer (PCA) (5). Therefore PSA is applied as a tumor marker for the clinical management of PCA (6). However, increased PSA concentrations in serum also occur in patients with benign prostate hyperplasia (BPH) (7). Hence the goal is to discriminate clearly between BPH and PCA in the clinical laboratory to spare the patient invasive diagnostic procedures, such as a prostate biopsy. In human serum PSA occurs in two forms: free PSA (f-PSA) and complexed PSA. The major form is a complex of PSA and a1-antichymotrypsin (ACT). The fraction of f-PSA was shown to be substantially smaller in patients with untreated PCA than in patients with BPH. Therefore combined measurements of f-PSA and total PSA (t-PSA) may lead to a better discrimination between BPH and PCA Some recent studies have already shown that the f-PSA/t-PSA ratio is 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 light. The employment of several serum references of known prostate specific antigen (fPSA) levels permits the construction of a dose response curve of activity and con_x001F_centration. From comparison to the dose response curve, an unknown specimen's activity can be correlated with fPSA concentration.