Growth Hormone 1 CLIA Kit

Catalog No. ABIN504794

This Chemiluminescent ELISA kit is designed for the quantitative measurement of Human Growth Hormone 1.

Quick Overview for Growth Hormone 1 CLIA Kit (ABIN504794)

Target: Growth Hormone 1 (GH1)

Reactivity: Human

Detection Method: Chemiluminescent

Method Type: Sandwich ELISA

Application: ELISA

Product Details Growth Hormone 1 CLIA Kit

Analytical Method: Quantitative

Characteristics: The Quantitative Determination of Growth Hormone Concentration in Human Serum by a Microplate Chemiluminescence Assay .

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 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_8(C 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_20(C for up to 30 days. Avoid repetitive freezing and thawing. When assayed in duplicate, 0.100 ml 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 microplate 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. Add 0.100 ml (100l) of hGH Tracer Reagent to all wells. 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 to all wells (see Reagent Preparation Section). ). Always add reagents in the same order to minimize reaction time differences between wells. Incubate for five (5) minutes in the dark. 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 substrate solution.

Restrictions: For Research Use only

Handling

Target Details

Target: Growth Hormone 1 (GH1)

Alternative Name: Growth hormone (Somatotropin)

Background: Growth hormone (hGH, somatotropin), secreted from the anterior pituitary, is a polypeptide with two intra-chain disulfide bridges, which circulates free or bound to number of different GH-binding proteins. Several forms of growth hormone have been identified (1) with the major being of molecular weight 22,000 daltons containing 191 amino acid residues. A 20,000-dalton variant, which posses all known biological functions of GH, has also been demonstrated to be important. The primary biological actions of the hormone are in direct growth promoting. GH exerts its effect directly on target organs such as bones and muscles and indirectly through the release of somatomedins, a family of insulin-like growth factor (IGF) hormones, produced in the liver (2). In particular, somatotropin C (IGF-1) is essential for bone growth during childhood. The clinical usefulness of the measurement of growth hormone (GH) in children has been well established in ascertaining linear bone growth along the epiphyseal plate. Abnormal elevated levels lead to gigantism while complete absence slows the rate of growth to one-third to one-half of normal. In adults, the epiphyseal growth plates have fused, GH excess gradually produces acromegaly, a coarse thickening of the bones of the skull, hands and feet. In this method, GH calibrator, patient specimen or control is first added to a streptavidin coated well. Biotinylated monoclonal and enzyme labeled antibodies (directed against distinct and different epitopes of GH) are added and the reactants mixed. Reaction between the various GH antibodies and native GH forms a sandwich complex that binds with the streptavidin coated to the well. After the completion of the required incubation period, the enzyme-growth hormone antibody bound conjugate is separated from the unbound enzyme-growth hormone 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 growth hormone levels permits the construction of a dose response curve of activity and concentration. From comparison to the dose response curve, an unknown specimen's activity can be correlated with growth hormone concentration.

Pathways: NF-kappaB Signaling, JAK-STAT Signaling, Intracellular Steroid Hormone Receptor Signaling Pathway, Peptide Hormone Metabolism, Regulation of Intracellular Steroid Hormone Receptor Signaling, Regulation of Hormone Metabolic Process, Response to Growth Hormone Stimulus, Regulation of Hormone Biosynthetic Process