Growth Hormone 1 Kit ELISA

N° du produit ABIN1305170

Détail du produit Growth Hormone 1 Kit ELISA

Antigène: Growth Hormone 1 (GH1)

Reactivité: Humain

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 0.087-24.0 ng/mL

Seuil minimal de détection: 0.087 ng/mL

Application: ELISA

Fonction: This ELISA (enzyme-linked immunosorbent assay) kit is intended forthe quantitative determination of human growth hormone (HGH) levelin serum. This assay is useful as an aid for diagnosis and treatment ofdisorders involving the anterior lobe of the pituitary gland.

Marque: ED™

Type d'échantillon: Serum

Analytical Method: Quantitative

Ingrédients: 1. Streptavidin Coated Microplate
Two vials each contain lyophilized human HGH in a bovine serum albumin-based matrix with a non-azide preservative. Refer to vials for exact concentration range for each control.

Both controls should be reconstituted with DI-water and store at -20 °C or below after the first use with up to 3 freeze-thaw cycles.

Matériel non inclus: 1. Precision single channel pipettes capable of delivering 25 µL, 50 µL, 100 µL, and 1000 µL etc.
2. Repeating dispenser suitable for delivering 100 µL.
3. Disposable pipette tips suitable for above volume dispensing.
4. Disposable 12 x 75 mm or 13 x 100 glass or plastic tubes.
5. Disposable plastic 100 mL and 1000 mL bottle with caps.
6. Aluminum foil.
7. Deionized or distilled water.
8. Plastic microtiter well cover or polyethylene film.
9. ELISA multichannel wash bottle or automatic (semi-automatic) washing system.
10. Spectrophotometric microplate reader capable of reading absorbance at 405 nm and 450 nm

Information d'application

Volume d'échantillon: 50 μL

Durée du test: 4 h

Plaque: Pre-coated

Protocole: This ELISA is designed, developed and produced for the quantitativemeasurement of human growth hormone in serum. The assay utilizesthe two-site sandwich technique with two selected HGH-specificmonoclonal antibodies that bind to different epitopes of human growthhormone.Assay standards, controls and patient serum samples containing HGHare added directly to microtiter wells of microplate that was coated withstreptavidin. Simultaneously, a biotinylated antibody and a horseradishperoxidase-conjugated antibody are added to each well. After the firstincubation period, the wall of microtiter well captures the biotinylatedantibody as well as an immunocomplex in the form of streptavidin biotin-antibody HGH HRP-antibody. Unbound proteins as well asunbound HRP conjugated antibody in each microtiter well are removedin the subsequent washing step. The well is incubated with a substratesolution in a timed reaction and then measured in a spectrophotometricmicroplate reader. The enzymatic activity of the tracer antibody boundto the HGH on the wall of the microtiter well is directly proportional tothe amount of HGH in the sample. A standard curve is generated byplotting the absorbance versus the respective HGH concentration foreach standard on Point-to-Point or 4-Parameter plot. Theconcentration of human HGH in test samples is determined directlyfrom this standard curve.

Préparation des réactifs:

(1) Prior to use allow all reagents to come to room temperature. Reagents from different kit lot numbers should not be combined or interchanged.
(2) ELISA Wash Concentrate must be diluted to working solution prior use. Please see REAGENTS section for details.
(3) Reconstitute all assay standards and controls by adding 0.5 mL of deionized water to each vial. Allow the standards and controls to sit undisturbed for 10 minutes, and then mix well by inversions or gentle vortexing. Make sure that all solids are dissolved completely prior to use. These reconstituted standards and controls must be stored at -10 C or below. Do not exceed 3 freeze-thaw cycles.
(4) Test Configuration
(5) Prepare working Tracer Antibody and Capture Antibody mixture by 1:21 fold dilution of the HGH Tracer Antibody and the HGH Capture Antibody with the Tracer Antibody Diluent . For each strip, it is required to mix 1 mL of Tracer Antibody Diluent with the addition of 50 µL of Tracer Antibody and 50 µL Capture Antibody in a clean test tube or vial. Following is a table that outlines the relationship of strips used and antibody mix prepared.

Prélèvement de l'échantillon: Only 50 µL of human serum is required for HGH measurement in duplicate. No special preparation of individual is necessary prior to specimen collection. Whole blood should be collected with Red-top Vaccutainer and separate the serum from cells by centrifugation (850 ? 1500xg for 10 minutes). Serum samples should be stored at -20 °C if the assay is not to be performed within 5 hours. Avoid more than three freeze-thaw cycles of specimen. Do not use grossly hemolyzed, icteric or lipemic samples.

Procédure de l'essai:

(1) Place a sufficient number of streptavidin coated microwell strips in a holder to run HGH standards, controls and unknown samples in duplicate.
(2) Add 25 µL of HGH standards, controls and patient serum samples into the designated microwell.
(3) Add 100 µL of above antibody mixture to each well.
(4) Mix gently and cover the plate with one plate sealer and also with aluminum foil to avoid exposure to light.
(5) Incubate plate at room temperature for 1 hour.
(6) Remove the aluminum foil and plate sealer. Aspirate the contents of each well. Wash each well 5 times by dispensing 350 µL of working wash solution into each well and then completely aspirating the contents. Alternatively, an automated microplate washer can be used.
(7) Add 100 µL of ELISA HRP Substrate into each of the wells.
(8) Cover the plate with one new plate sealer and also with aluminum foil to avoid exposure to light.
(9) Incubate plate at room temperature for 15 minutes. (This incubation period may be reduced to 8 ? 10 min if a lower OD reading is demanded to fit to the plate readers specification.)
(10) Remove the foil and plate sealer. Add 100 µL of ELISA Stop Solution into each of the wells. Mix gently.
(11) Read the absorbance at 405 nm and 450 nm within 10 minutes in a microplate reader.

Calcul des résultats:

1. Calculate the average absorbance for each pair of duplicate test results.
   2. Subtract the average absorbance of the STD 1 (0 ng/mL) from the average absorbance of all other readings to obtain corrected absorbance.
   3. The standard curve is generated by the corrected absorbance of all standard levels on the ordinate against the standard concentration on the abscissa using point-to-point or log-log paper. Appropriate computer assisted data reduction programs may also be used for the calculation of results.

Précision du teste: The intra-assay precision is validated by measuring two controls samples in a single assay with 20 replicate determinations. The inter-assay precision is validated by measuring two control samples in duplicate in 12 individual assays.

Restrictions: For Research Use only

Stockage

Précaution d'utilisation: The reagents must be used in a professional laboratory environment and are for in vitro diagnostic use. The source material for reagents containing bovine serum was derived in the contiguous 48 United States. It was obtained only from healthy donor animals maintained under veterinary supervision and found free of contagious diseases. Wear gloves while performing this assay and handle these reagents as if they were potentially infectious. Avoid contact with reagents containing TMB, hydrogen peroxide, or sulfuric acid. TMB may cause irritation to skin and mucous membranes and cause an allergic skin reaction. TMB is a suspected carcinogen. Hydroxide chloride may cause severe irritation and damage on contact with skin. Do not get in eyes, on skin, or on clothing. Do not ingest or inhale fumes. On contact, flush with copious amounts of water for at least 15 minutes. Wear laboratory gloves, eye glasses, etc. Use Good Laboratory Practices.

Stock: 4 °C

Détail du antigène

Antigène: Growth Hormone 1 (GH1)

Autre désignation: Growth Hormone (GH1 Produits)

Synonymes: GH Kit ELISA, GH-N Kit ELISA, GHN Kit ELISA, IGHD1B Kit ELISA, hGH-N Kit ELISA, Gh1 Kit ELISA, GH1 Kit ELISA, gh Kit ELISA, Gh Kit ELISA, GH2 Kit ELISA, RNGHGP Kit ELISA, ghl Kit ELISA, gh-n Kit ELISA, ghn Kit ELISA, ighd1b Kit ELISA, gh1 Kit ELISA, ghb-A Kit ELISA, GHI Kit ELISA, GHB3 Kit ELISA, growth hormone 1 Kit ELISA, growth hormone Kit ELISA, somatotropin-like Kit ELISA, somatotropin Kit ELISA, growth hormone prepeptide Kit ELISA, growth hormone 1 L homeolog Kit ELISA, growth hormone 1 S homeolog Kit ELISA, Somatotropin-1 Kit ELISA, GH1 Kit ELISA, Gh Kit ELISA, GH Kit ELISA, LOC100305005 Kit ELISA, Gh1 Kit ELISA, LOC100534452 Kit ELISA, LOC100232594 Kit ELISA, LOC100303681 Kit ELISA, gh1 Kit ELISA, LOC100356068 Kit ELISA, LOC100136588 Kit ELISA, gh1.L Kit ELISA, gh1.S Kit ELISA, LOC109081196 Kit ELISA

Classe de substances: Hormone

Pathways: Signalisation NF-kappaB, Signalistation JAK/STAT, 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