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Progesterone CLIA Kit

Reactivity: Human Chemiluminescent Competition ELISA
Catalog No. ABIN504754
  • Target See all Progesterone CLIA Kits
    Progesterone
    Reactivity
    • 4
    • 3
    • 3
    • 3
    • 3
    • 3
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Human
    Detection Method
    Chemiluminescent
    Method Type
    Competition ELISA
    Application
    ELISA
    Purpose
    Competitive Enzyme Immunoassay (TYPE 7): The essential reagents required for a 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 Progesterone Concentration in Human Serum or Plasma by a Microplate Chemiluminescence Immunoassay
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  • 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 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 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 0.025 ml (25 L) of the appropriate serum reference, control or specimen into the assigned well. 3. Add 0.050 ml (50l) of the Progesterone Tracer Reagent to all wells. 4. Swirl the microplate gently for 20-30 seconds to mix. 5. Add 0.050 ml (50l) of the Progesterone Biotin Reagent to all wells. 6. Swirl the microplate gently for 20-30 seconds to mix. 7. Cover and incubate for 60 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 samples suspected of concentrations higher than 60ng/ml 1:5 and 1:10 with progesterone 0 ng/ml calibrator or male patient serum pools with a known low value for progesterone.
    Restrictions
    For Research Use only
  • Target See all Progesterone CLIA Kits
    Progesterone
    Abstract
    Progesterone Products
    Target Type
    Hormone
    Background
    Measurement of progesterone in serum or plasma is considered to be the most reliable way to assess its rate of production. Progesterone is a steroid hormone, which plays an important role in the preparation for and maintenance of pregnancy. It is synthesized from cholesterol via pregnenolone then rapidly metabolized to pregnanediol primarily in the liver 2, 9, 13. The ovary and placenta are the major production sites, but a small amount is also produced by the adrenal cortex in both men and women. Circulating progesterone levels, which are characteristically low during the follicular phase, increase sharply during the luteal phase of menstrual cycles, reaching a maximum approximately 5 to 10 days after the midcycle LH peak12. Unless pregnancy occurs, a steep decline to follicular levels sets in about 4 days before the next menstrual period. This pattern constitutes the rationale behind the well established use of serum progesterone measurements as a simple and reliable method for ovulation detection3, 4, 16. For routine measurements, immunoassays using steroid specific antibodies are preferred. Initial immunoassays, for serum progesterone, used organic solvents to remove the steroid from endogenous binding proteins such as corticosteroid binding globulin (CBG) and albumin. Direct measurement of progesterone in serum or plasma is considered to be the method of choice for routine applications. Both RIA and EIA (and some FIA) are available in the market. Since RIA involves handling radioactivity and causes radioactive waste disposal issues, various non-isotopic methods have replaced the RIA. These methods use very specific antibodies to determine levels of progesterone in circulation. The Monobind Progesterone CLIA kit uses a specific anti-progesterone antibody, and does not require 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 progesterone concentration permits construction of a graph of activity and con_x001F_centration. From comparison to the dose response curve, an unknown specimen's activity can be correlated with progesterone concentration.
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