Chromogranin A Kit ELISA

N° du produit ABIN1305163

Détail du produit Chromogranin A Kit ELISA

Antigène: Chromogranin A (CHGA)

Reactivité: Humain

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 2.0-660 ng/mL

Seuil minimal de détection: 2 ng/mL

Application: ELISA

Fonction: This ELISA (enzyme-linked immunosorbent assay) kit is intended for the quantitative determination of human chromogranin A levels in EDTA-plasma and serum samples.

Marque: ED™

Type d'échantillon: Serum, Plasma

Analytical Method: Quantitative

Ingrédients: Anti-CgA Antibody Coated Microplate , Manual, Certificate of Analysis

Matériel non inclus: 1. Precision single channel pipettes capable of delivering 15, µL, 50 µL, 100 µL, and 1000 µL etc., Repeating dispenser suitable for delivering 100 µL., Disposable pipette tips suitable for above volume dispensing., Disposable 12 x 75 mm or 13 x 100 glass or plastic tubes., Disposable plastic 100 mL and 1000 mL bottle with caps., Aluminum foil., Deionized water., Plastic microtiter well cover or polyethylene film., ELISA multichannel wash bottle or automatic (semi-automatic) washing system. Spectrophotometric microplate reader capable of reading absorbance at 450/650 nm or 450/620 nm.

Information d'application

Volume d'échantillon: 15 μL

Durée du test: 4 h

Plaque: Pre-coated

Protocole: Assay standards, controls and patient samples are added directly to wells of microplate that is coated with a polyclonal chromogranin A antibody. After the first incubation period, the antibody on the wall of microtiter well captures human chromogranin A in the sample and unbound protein in each microtiter well is washed away. Then a horseradish peroxidase (HRP)-labeled monoclonal anti-human chromogranin A antibody is added to each microtiter well and a sandwich of monoclonal antibody - human chromogranin A polyclonal antibody is formed. The unbound monoclonal antibody is removed in the subsequent washing step. For the detection of this immunocomplex, the well is incubated with a substrate solution in a timed reaction and then measured in a spectrophotometric microplate reader. The enzymatic activity of the immunocomplex bound to the chromogranin A on the wall of the microtiter well is directly proportional to the amount of chromogranin A in the sample. A standard curve is generated by plotting the absorbance versus the respective human chromogranin A concentration for each standard with a 4 parameter curve fit. The concentration of human chromogranin A in test samples is determined directly from 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 deminerialized 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.

Prélèvement de l'échantillon: Only 30 µL total (15 µL each) of human EDTA-plasma or serum is required for human chromogranin A measurement in duplicate. No special preparation of individual is necessary prior to specimen collection. Whole blood should be collected with lavender-top Vaccutainer. Separate the plasma from cells by centrifugation (850 ? 1500xg for 10 minutes). The plasma should be separated from the cells within one hour of blood collection and transferred to a clean test tube. Plasma samples should be stored at ? 15 C if the assay is not to be performed within 72 hours. Otherwise, the plasma samples should be stored at room temperature for up to 72 hours. It is important that the plasma samples must not be stored at 2 ?8 C for long-term storage. Avoid more than three freeze-thaw cycles of specimen. Serum sample can also be used for chromogranin A measurement. Tests with paired EDTA-plasma and serum sample from same donor shows that serum gives almost the same chromogranin A level as EDTA-plasma by using this ELISA.

Procédure de l'essai:

(1) Place a sufficient number of antibody coated microwell strips in a holder to run human chromogranin A standards, controls and unknown samples in duplicate.
(2) Add 15 µL of standards, controls and patient samples into the designated microwells.
(3) Add 200 µL of assay buffer to each well.
(4) Cover the plate with one plate sealer and also with aluminum foil, and incubate plate on an ELISA plate shaker with a shaking rate at 350 rpm to 450 rpm at room temperature for 1 hour.
(5) Remove 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.
(6) Add 100 µL of Chromogranin A Tracer Antibody to each of the wells.
(7) Cover the plate with the plate sealer and also with aluminum foil, and incubate plate on an ELISA plate shaker with a shaking rate at 350 rpm to 450 rpm at room temperature for 1 hour.
(8) Remove 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.
(9) Add 100 µL of ELISA HRP Substrate into each of the wells.
(10) Cover the plate with one plate sealer and also with aluminum foil to avoid exposure to light.
(11) Incubate plate at room temperature for 20 minutes.
(12) Remove the aluminum foil and plate sealer. Add 100 µL of ELISA Stop Solution into each of the wells. Mix gently.(13) Read the absorbance at dual wavelength at 450/650 nm or 450/620 nm within 10 minutes in a microplate reader and select a 4-parameter curve fit for result calculation.

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. The human chromogranin A concentrations for the controls and patient samples are read directly from the standard curve using their respective corrected absorbanceTo assure the validity of the results each assay should include adequate controls with known chromogranin A levels. We recommend that all assays include the laboratory's own chromogranin A controls in addition to those provided with this kit.

Précision du teste: The inter-assay precision is validated by measuring two control samples in duplicate in 12 individual assays. The intra-assay precision is validated by measuring two controls samples in a single assay with 8 replicate determinations.

Restrictions: For Research Use only

Stockage

Précaution d'utilisation: The reagents must be used in a professional laboratory environment and are for research use only. 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. Sulfuric acid may cause severe irritation 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. Use Good Laboratory Practices.

Stock: 4 °C

Détail du antigène

Antigène: Chromogranin A (CHGA)

Autre désignation: Chromogranin A (CHGA Produits)

Synonymes: CGA Kit ELISA, CG-ALPHA Kit ELISA, FSHA Kit ELISA, GPHA1 Kit ELISA, GPHa Kit ELISA, HCG Kit ELISA, LHA Kit ELISA, TSHA Kit ELISA, ChrA Kit ELISA, fj05f09 Kit ELISA, si:dkey-177p2.2 Kit ELISA, wu:fj05f09 Kit ELISA, zgc:101749 Kit ELISA, zgc:56075 Kit ELISA, CHGA Kit ELISA, cgA Kit ELISA, chga Kit ELISA, chromogranin A Kit ELISA, glycoprotein hormones, alpha polypeptide Kit ELISA, uncharacterized CHGA Kit ELISA, chromogranin A S homeolog Kit ELISA, CHGA Kit ELISA, CGA Kit ELISA, Chga Kit ELISA, chga Kit ELISA, chga.S Kit ELISA

Sujet: Chromogranin A is a 49 kDa acidic protein that consists of 439 amino acids encoded on chromosome 14. Chromogranin A has been identified in a number of normal and neopastic endocrine tissues. It is demonstrated that an elevated level of circulating chromogranin A is a marker for tumors of neuroendocrine origin. However, the most significant clinical use of chromogranin A is related to the diagnostic procedure in patients with pheochromocytoma. The following is a short summary of the potential usages of chromogranin A.1. A very sensitive (83 % ) and highly specific (96 %) marker in the evaluation of actual or suspected pheochromocytoma. Drugs commonly employed in the diagnosis or treatment of pheochromocytoma have little effect on the plasma chromogranin A level, which is a great advantage of measuring chromogranin A over catecholamines. 2. To ascertain the source of a tumor. A high chromogranin A level indicates that the tumor arises from neuroendocrine tissues.3. Endocrine tumors that do not produce their specific hormones, for example, calcitonin negative but chromogranin A positive C-cell carcinoma, zero-cell carcinoma, beta-cell carcinoma, parathyroid carcinoma.

Poids moléculaire: 49 kDa

ID gène: 1113

NCBI Accession: NP_001266

UniProt: P10645

Pathways: Negative Regulation of Hormone Secretion, cAMP Metabolic Process, Regulation of G-Protein Coupled Receptor Protein Signaling