Cortisol Kit CLIA

N° du produit ABIN504753

Détail du produit Cortisol Kit CLIA

Antigène: Cortisol

Reactivité: Humain

Méthode de détection: Chemiluminescent

Type de méthode: Competition ELISA

Application: ELISA

Fonction: Competitive Enzyme Immunoassay (TYPE 7): The essential reagents required for a chemiluminescence 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

Attributs du produit: The Quantitative Determination of Total Cortisol Concentration in Human Serum or Plasma 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: 10 μL

Plaque: Pre-coated

Protocole:

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, 0.050ml of the specimen is required.

Reagent Preparation:

1. Working Tracer Reagent (Stable for 1 year) Dispense 0.7 ml of Cortisol Tracer Reagent and add to the vial containing Steroid Tracer Buffer. Store at 2-8C. 2. 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. 3. 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 reaction 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 serum reference, control or specimen into the assigned well. 3. Add 0.050 ml (50l) of the working Cortisol Tracer Reagent to all wells (see Reagent Preparation Section). 4. Swirl the microplate gently for 20-30 seconds to mix. 5. Add 0.050 ml (50l) of Cortisol 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 substrate. Note: Dilute the samples suspected of concentrations higher than 50 g/dl 1:5 and 1:10 with cortisol 0 g/dl patient serum.

Restrictions: For Research Use only

Stockage

Détail du antigène

Antigène: Cortisol

Abstract: Cortisol Produits

Classe de substances: Hormone

Sujet: Cortisol (hydrocortisone, compound F) is the most potent glucocorticoid produced by the human adrenal cortex. As with other adrenal steroids, cortisol is synthesized from cholesterol, through a series of enzymatically mediated steps, by the adrenal cortex [reviewed in 1, 2]. The first and rate-limiting step in adrenal steroidogenesis, conversion of cholesterol to pregnenolone, is stimulated by pituitary adrenocorticotropic hormone (ACTH) which is, in turn, regulated by hypothalamic corticotropin releasing factor (CRF). ACTH and CRF secretion are inhibited by high cortisol levels. In plasma, the major portion of cortisol is bound with high affinity to corticosteroid-binding globulin (CBG, transcortin), with most of the remainder loosely bound to albumin. Physiologically effective in anti-inflammatory activity and blood pressure maintenance, cortisol is also involved in gluconeogenesis. Cortisol acts through specific intracellular receptors and has effects in numerous other physiologic systems, including immune function, glucose-counter regulation, vascular tone, substrate utilization and bone metabolism [1-3]. Cortisol is excreted primarily in urine in an unbound (free) form. Cortisol production has an ACTH-dependent circadian rhythm with peak levels in the early morning and a nadir at night. The factors controlling this circadian rhythm are not completely defined. The circadian rhythm of ACTH/cortisol secretion matures gradually during early infancy, and is disrupted in a number of physical and psychological conditions [4]. Furthermore, increased amounts of ACTH and cortisol are secreted independently of the circadian rhythm in response to physical and psychological stress [4, 5]. Elevated cortisol levels and lack of diurnal variation have been identified in patients with Cushing's disease (ACTH hyper secretion) [2, 6]. Elevated circulating cortisol levels have also been identified in patients with adrenal tumors [7]. Low cortisol levels are found in primary adrenal insufficiency (e.g. adrenal hypoplasia, congenital adrenal hyperplasia, Addison's disease) and in ACTH deficiency [1, 2, 8, and 9]. Due to the normal circadian variation of cortisol levels, distinguishing normal and abnormally low cortisol levels can be difficult. Therefore, various tests to evaluate the pituitary-adrenal (ACTH-cortisol) axis, including insulin-induced hypoglycemia, short- and long-term ACTH stimulation, CRF stimulation and artificial blockage of cortisol synthesis with metronome have been performed [8-10]. Cortisol response characteristics for each of these procedures have been reported. The Monobind Cortisol CLIA Kit uses a specific monoclonal anti-cortisol antibody, and does not require prior sample extraction of serum or plasma. Cross-reactivity to other naturally-occurring steroids is low. The employment of several serum references of known cortisol 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 cortisol concentration.