CEA, AFP, PSA CLIA Kit

Catalog No. ABIN504768

The Human CEA, AFP, PSA ELISA Kit (ABIN504768) is a Chemiluminescent ELISA Kit designed to quantify Human CEA, AFP, PSA.

Quick Overview for CEA, AFP, PSA CLIA Kit (ABIN504768)

Target: CEA, AFP, PSA

Reactivity: Human

Detection Method: Chemiluminescent

Method Type: Sandwich ELISA

Application: ELISA

Product Details

Purpose: 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-marker specific 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

Characteristics: The Quantitative Determination of AFP, CEA and PSA Concentration in Human Serum and Plasma by a Chemiluminescence Immunoassay (CLIA)). Measurements of these tumor markers are used as an aid in the diagnosis and monitoring of various oncological disorders.

Application Details

Application Notes: 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.

Sample Volume: 25 μL

Plate: Pre-coated

Protocol:

Specimien Collection and Preparation:

The specimens shall be blood, serum or plasma 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 (for serum) or evacuated tube(s) containing EDTA or heparin.. Allow the blood to clot for serum samples. Centrifuge the specimen to separate the serum or plasma from the cells. Samples may be refrigerated at 2_x001E_8oC 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_20oC for up to 30 days. Avoid repetitive freezing and thawing.. When assayed in duplicate, 0.05 ml of the specimen is required for each tumor marker assayed.

Reagent Preparation:

1. Wash Buffer Dilute contents of Wash Concentrate to 1000ml with distilled or deionized water in a suitable storage container. Store at room temperature until expiration date printed on concentrate label. 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 0.025ml (25l) of the appropriate serum reference, control or specimen into the assigned well. 3. Add 0.100ml (100l) of the appropriate tracer reagent to each well. It is very important to use the right Tracer Reagent for each assay for accurate results. 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 0.100ml (100l) of working signal reagent solution to all wells (see Reagent Preparation Section). Always add reagents in the same order to minimize reaction time differences between wells. 9. Incubate for five (5) minutes at room temperature in the dark. 10. Read the RLUs (Relative Light Units) in each well in a microplate luminometer for at least 0.2 seconds/ well. The results can be read within 30 minutes of adding the substrate solution. Note: It is very important to dispense all reagents in the center of the coated well. Always add reagents in the same order to minimize reaction time differences between wells. _x000C_

Restrictions: For Research Use only

Handling

Target Details

Target: CEA, AFP, PSA

Background: Alpha-Fetoprotein (AFP) is a glycoprotein with a molecular weight of 70 kDA. AFP is normally produced during fetal development by the hepatocytes, yolk sac and to a lesser extent by the gastrointestinal tract. Serum concentrations reach a peak level of up to 10 mg/ml at twelve weeks of gestation (1). This peak level gradually decreases to less than 25 ng/ml after one year of postpartum. Thereafter, the levels reduce further to less than 10 ng/ml. Elevated levels of AFP are found in patients with primary heptatoma and yolk sac-derived germ tumors. AFP is the most useful marker for the diagnosis and management of hepatocellular carcinoma (2). AFP is also elevated in pregnant women. Presence of abnormally high AFP concentrations in pregnant women provides a risk marker for Down syndrome (3), Carcinoembryonic antigen (CEA) is a glycoprotein with a molecular weight of 180 kDA. CEA is the first of the so-called carcinoembryonic proteins that was discovered in 1965 by Gold and Freeman (5). CEA is the most widely used marker for gastrointestinal cancer. Although CEA is primarily associated with colorectal cancers, other malignancies that can cause elevated levels of CEA include breast, lung, stomach, pancreas, ovary and other organs. Benign conditions that cause significantly higher than normal levels include inflammation of lung and gastrointestinal (GI) tract and benign liver cancer (6,8). Heavy Smokers, as a group, have higher than normal baseline concentration of CEA. Prostate Specific antigen (PSA) is a serine protease with chymotrypsin-like activity (15, 17, and 19). The protein is a single chain glycoprotein with a molecular weight of 28.4 kDA (16). 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 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.