Estriol CLIA Kit

Catalog No. ABIN504758

This Chemiluminescent ELISA kit is designed for the quantitative measurement of Human Estriol.

Quick Overview for Estriol CLIA Kit (ABIN504758)

Target: Estriol

Reactivity: Human

Detection Method: Chemiluminescent

Method Type: Competition ELISA

Application: ELISA

Product Details Estriol CLIA Kit

Purpose: Competitive Enzyme Immunoassay (TYPE 7): The essential reagents required for an enzyme immunoassay include antibody, enzyme-antigen conjugate and native antigen. Upon mixing biotinylated antibody, enzyme-antigen conjugate and a serum containing the native antigen, a competition reaction results between the native antigen and the enzyme-antigen conjugate for a limited number of antibody binding sites.

Analytical Method: Quantitative

Characteristics: The Quantitative Determination of Free (unconjugated) Estriol (u-E3) Concentration in Human Serum or Plasma by a Microplate Chemiluminescence Immunoassay

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.

Plate: Pre-coated

Protocol:

Specimien Collection and Preparation:

The specimens shall be blood, serum or heparanised plasma in type and taken with the usual precautions in the collection of venipuncture samples. The blood should be collected in a redtop (with or without gel additives) venipuncture tube or for plasma use evacuated tube(s) containing 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, 0.050ml 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 for up to 60 days. 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 1 ml 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 0.025 ml (25l) of the appropriate calibrator, control or specimen into the assigned well. 3. Add 0.050 ml (50l) of the u-Estriol Tracer Reagent to all wells. 4. Swirl the microplate gently for 20-30 seconds to mix. 5. Add 0.050 ml (50l) of u-Estriol Biotin Reagent to all wells. 6. Swirl the microplate gently for 20-30 seconds to mix. 7. Cover and incubate for 45 minutes at room temperature. 8. Discard the contents of the microplate by decantation or aspiration. If decanting, blot the plate dry with absorbent paper. 9 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. 10. Add 0.100 ml (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. 11. Incubate at room temperature for five (5) minutes in the dark. 12. Read the relative light units in each well with a Chemiluminescence microplate reader for 0.5-1.0 seconds. The results should be read within 30 minutes after adding the working Signal Reagent. Note: Dilute the sample, suspected of concentrations higher than 30ng/ml, by diluting 1:2 and/or 1:5 with unconjugated estriol 0 ng/ml calibrator or male patient sera with a known low value for estriol. Multiply the result by the dilution factor of 2 or 5 as required to obtain the concentration of the sample.

Restrictions: For Research Use only

Handling

Target Details

Target: Estriol

Target Type: Hormone

Background: The last few years have seen the development of screening for fetal Down syndrome by measurement of multiple markers in maternal blood (1). Although amniocentesis has been widely available for more than 40 years it can only be selectively used to diagnose the disorder because of the hazard to fetus. Of most employed for differential diagnosis the commonly used procedures are AFP, hCG, free beta-HCG and unconjugated estriol(2). Unconjugated estriol in the serum of pregnant women originates almost exclusively from precursors in the fetus, via the placenta.(3) The clinical evidence shows that in uncomplicated pregnancies, the production of estriol increases steadily throughout the last trimester, however, in pregnancies complicated by placental insufficiency the synthesis of estriol decreases rapidly. For many years the most commonly used method for monitoring estriol synthesis (as an index to fetal stress) has been to measure estriol and estriol conjugates in a 24 hr urine sample(4). However, changes in renal clearance and diurnal variations can make the results of these determinations suspect. In recent years investigators have found the determinations of unconjugated estriol in pregnancy plasma as an alternative to the urinary assay to be a better marker of fetal stress(6). Abnormally low levels of estriol in a pregnant woman may indicate a problem with the development in the child. Levels of estriol in non-pregnant women do not change much after menopause, and levels are not significantly different from levels in men (7) The Monobind unconjugated estriol CLIA kit uses a specific anti-estriol antibody, and does not require prior sample extraction of serum or plasma. Cross-reactivity to other naturally-occurring and structurally related steroids is low. The employment of several serum references of known Estriol concentration permits construction of a graph of activity (light) and con_x001F_centration. From comparison to the dose response curve, an unknown specimen's activity can be correlated with Estriol concentration.