AMH Kit ELISA

N° du produit ABIN3073425

Détail du produit AMH Kit ELISA

Antigène: AMH (Anti-Mullerian Hormone (AMH))

Reactivité: Humain, Non-Human Primate

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 0.084-14.2 ng/mL

Seuil minimal de détection: 0.084 ng/mL

Application: ELISA

Fonction: The Ultra-Sensitive Anti-Müllerian hormone/Müllerian inhibiting substance (US AMH/MIS) enzyme linked immunuosorbent assay (ELISA) kit provides materials for the quantitative measurement of AMH/MIS in human serum, plasma and other biological fluids. This kit is intended for laboratory research use only and is not for use in diagnostic or therapeutic procedures.

Type d'échantillon: Plasma, Serum

Analytical Method: Quantitative

Sensibilité: 23 pg/mL

Ingrédients:

• AMH/MIS Calibrators A / Sample Diluent
• AMH/MIS Calibrators B thru F (Lyophilized)
• AMH/MIS Controls I & II (Lyophilized)
• AMH/MIS Coated Microtitration strip
• AMH/MIS Assay Buffer
• AMH Biotin Conjugate Ready-To-Use (RTU)
• AMH/MIS Streptavidin-Enzyme Conjugate-Ready-to-Use (RTU)
• TMB Chromogen Solution
• Stopping Solution
• Wash Concentrate A

Matériel non inclus:

1. Microtitration plate reader capable of absorbance measurement at 450 nm, 405nm and 630 nm.
2. Microplate orbital shaker.
3. Microplate washer.
4. Semi-automated/manual precision pipette to deliver 10-250 μL.
5. Repeator pipette.
6. Vortex mixer.
7. Deionized water.

Information d'application

Volume d'échantillon: 25 μL

Durée du test: 2.5 h

Plaque: Pre-coated

Préparation des réactifs:

1. AMH/MIS Calibrators B-F and AMH/MIS Controls I & II: Tap and reconstitute AMH/MIS Calibrator B-F and AMH/MIS Controls I & II each with 1 mL deionized water. Solubilize, mix well and use after reconstitution.
2. Wash Solution: Dilute wash concentrate 25-fold with deionized water. The wash solution is stable for one month at room temperature (23 ± 2 °C) when stored in a tightly sealed bottle.
3. Microtitration Wells: Select the number of coated wells required for the assay. The remaining unused wells should be placed in the resealable pouch with a desiccant. The pouch must be resealed to protect from moisture.

Prélèvement de l'échantillon:

• Serum and lithium-heparin plasma is the recommended sample type.
• Sample handling, processing, and storage requirements depend on the brand of blood collection tube that you use. Please reference the manufacturer's instructions for guidance. Each laboratory should determine the acceptability of its own blood collection tubes and serum separation products.
• Samples may be stored at 4 °C if assayed within 24 hours, otherwise samples must be stored at -20 °C or -80 °C to avoid loss of bioactivity and contamination.
• Avoid assaying lipemic, hemolyzed or icteric samples.
• Avoid repeated freezing and thawing of samples. Thaw samples no more than 3 times.
• For shipping, place specimens in leak proof containers in biohazard specimen bags with appropriate specimen identification and test requisition information in the outside pocket of the biohazard specimen bag. Follow DOT and IATA requirements when shipping specimens.

Procédure de l'essai:

Allow all specimens and reagents to reach room temperature (23 ± 2 °C) and mix thoroughly by gentle inversion before use. Calibrators, controls, and unknowns should be assayed in duplicate. NOTE: All serum samples reading higher than the highest calibrator should be mixed and diluted in the 0 ng/mL Calibrator A/Sample diluent prior to assay.

1. Reconstitute AMH/MIS Calibrator B-F and AMH/MIS Controls I & II each with 1 mL deionized water. Solubilize for 10 minutes, Mix well by gentle vortex.
2. Label the microtitration strips to be used.
3. Pipette 25 μL of the Calibrator, Controls and Unknowns to the appropriate wells.
4. Add 100 μL of the AMH/MIS Assay Buffer to each well using a repeater pipette.
5. Incubate the plate, shaking at a fast speed (600-800 rpm) on an orbital microplate shaker, for 90 minutes at room temperature (23 ± 2 °C).
6. Aspirate and wash each strip 5 times (350 μL/per well) with Wash Solution using an automatic microplate washer.
7. Add 100 μL of the Antibody-Biotin Conjugate RTU to each well using a repeater pipette.
8. Incubate the plate, shaking at a fast speed (600-800 rpm) on an orbital microplate shaker, for 30 minutes at room temperature (23 ± 2 °C).
9. Aspirate and wash each strip 5 times with the Wash Solution (350 μL/per well) using an automatic microplate washer.
10. Add 100 μL of the Streptavidin-Enzyme Conjugate-RTU to each well using a repeater pipette.
11. Incubate the plate, shaking at a fast speed (600-800 rpm) on an orbital microplate shaker, for 30 minutes at room temperature (23 ± 2 °C).
12. Aspirate and wash each strip 5 times with the Wash Solution (350 μL/per well) using an automatic microplate washer.
13. Add 100 μL of the TMB chromogen solution to each well using a repeater pipette. Avoid exposure to direct sunlight.
14. Incubate the wells, shaking at 600-800 rpm on an orbital microplate shaker, for 8-12 min at room temperature (23 ± 2 °C). NOTE: Visually monitor the color development to optimize the incubation time.
15. Add 100 μL of the stopping solution to each well using a repeater pipette. Read the absorbance of the solution in the wells within 20 minutes, using a microplate reader set to 450 nm. NOTE: Zero calibrator should be programmed as "Blank" while reading the optical density. If instrument has a wavelength correction, set the instrument to dual wavelength measurement at 450 nm with background wavelength correction at 630 nm.

Calcul des résultats:

NOTE: The results in this package insert were calculated by plotting the log optical density (OD) data on the y-axis and log AMH concentration on X- axis using a cubic regression curve-fit. Alternatively, log vs. log quadratic regression curve-fit can be used. Other data reduction methods may give slightly different results.

1. Optimum results can be obtained at incubation temperature of (23 ± 2 °C).
2. Calculate the mean optical density (OD) for each calibrator, Control, or Unknown.
3. Plot the log of the mean OD readings for each of the Calibrators along the y-axis versus log of the AMH/MIS concentrations in ng/mL along the x-axis, using a cubic regression curve-fit.
4. Determine the AMH/MIS concentrations of the Controls and unknowns from the calibration curve by matching their mean OD readings with the corresponding AMH/MIS concentrations.
5. Any sample reading higher than the highest Calibrator should be appropriately diluted with the 0 ng/mL (CAL A / Sample Diluent) and re- assayed.
6. Any sample reading lower than the analytical sensitivity should be reported as such.
7. Multiply the value by a dilution factor, if required.

Précision du teste: Reproducibility of the US AMH/MIS ELISA assay was determined using three serum pools for a total of 12 assays. At concentrations between 0.3 and 1.8 ng/mL, the CV's ranged between 4.4 and 6.03%.

Restrictions: For Research Use only

Stockage

Précaution d'utilisation: For Research Use Only. Not for use in diagnostic procedures. The following precautions should be observed: a) Follow good laboratory practice. b) Use personal protective equipment. Wear lab coats and disposable gloves when handling immunoassay materials. c) Handle and dispose of all reagents and material in compliance with applicable regulations. WARNING: Potential Biohazardous Material This reagent may contain some human source material (e.g. serum) or materials used in conjunction with human source materials. Handle all reagents and patient samples at a Biosafety Level 2, as recommended for any potentially infectious human material in the Centers for Disease Control/National Institutes of Health manual "Biosafety in Microbiological and Biomedical Laboratories," 5th Edition, 2007. WARNING: Potential Chemical Hazard Some reagents in this kit contain Pro-Clean 400 and Sodium azide4 as a preservative. Pro-Clean 400 and Sodium azide in concentrated amounts are irritants to skin and mucous membranes. For further information regarding hazardous substances in the kit, please refer to the MSDS.

Stock: 4 °C

Détail du antigène

Antigène: AMH (Anti-Mullerian Hormone (AMH))

Autre désignation: AMH (AMH Produits)

Synonymes: AMH Kit ELISA, amh Kit ELISA, MIF Kit ELISA, MIS Kit ELISA, anti-Mullerian hormone Kit ELISA, amh Kit ELISA, AMH Kit ELISA, Amh Kit ELISA

Pathways: Negative Regulation of Hormone Secretion