AMH ELISA Kit

Produktnummer ABIN5680711

Produktdetails AMH ELISA Kit

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

Reaktivität: Schwein

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 0.2 ng/mL - 13 ng/mL

Untere Nachweisgrenze: 0.2 ng/mL

Applikation: ELISA

Verwendungszweck: The Porcine Anti-Müllerian hormone (AMH) enzyme-linked immunosorbent assay (ELISA) kit provides materials for the quantitative measurement of AMH in porcine serum and other biological fluids.

Proben: Serum

Analytische Methode: Quantitative

Sensitivität: 0.055 ng/mL

Bestandteile:

• AMH/MIS Calibrators A / Sample Diluent
• Canine AMH Calibrator H (Lyophilized)
• AMH Coated Microtitration strips
• AMH Assay Buffer
• AMH Biotin Conjugate Ready-To-Use (RTU)
• AMH Streptavidin-Enzyme Conjugate-Ready-to-Use (RTU)
• TMB Chromogen Solution
• Stopping Solution
• Wash Concentrate A

Benötigtes Material:

1. Microtitration plate reader capable of absorbance measurement at 450 nm, 405nm and 630 nm.
2. Microplate shaker.
3. Microplate washer.
4. Semi-automated/manual precision pipette to deliver 10-250 μL.
5. Disposable 12 x 75 mm culture tubes.
6. Vortex mixer.
7. Deionized water.

Anwendungsinformationen

Probenmenge: 50 μL

Testdauer: 3.5 h

Plattentyp: Pre-coated

Aufbereitung der Reagenzien:

1. AMH Calibrator H: a. Tap and reconstitute AMH Calibrator H with 1 mL deionized water. Solubilize for ten minutes, mix well before use. b. Prepare seven polystyrene tubes and label them as Cal A, Cal B, Cal C, Cal D, Cal E, Cal F and Cal G. c. Add 500 μL of AMH/MIS Calibrator A/Sample Diluent to each polystyrene tube labeled Cal A-G. d. Add 500 μL of reconstituted AMH Calibrator H (from step a) to the tube labeled Cal G. Vortex and mix the content in the tube thoroughly before the next dilution transfer. e. Add 500 μLof Cal G (from step d) to the tube labeled Cal F. Vortex and mix the content in the tube thoroughly before the next dilution transfer. f. Add 500 μL of Cal F (from step e) to the tube labeled Cal E. Vortex and mix the content in the tube thoroughly before the next dilution transfer. g. Add 500 μL of Cal E (from step f) to the tube labeled Cal D. Vortex and mix the content in the tube thoroughly before use. h. Add 500 μL of Cal D (from step g) to the tube labeled Cal C. Vortex and mix the content in the tube thoroughly before use. i. Add 500 μL of Cal C (from step h) to the tube labeled Cal B. Vortex and mix the content in the tube thoroughly before use. Note: If more sensitivity is desired, Cal B can be further diluted to B/2 and B/4 using Cal A. j. The tube labeled Cal A contains 500 μL AMH Calibrator A/Sample Diluent and has 0 AMH concentrations and should be used as Blank. k. The Calibrators A-G for instance should read as 0.0, 0.39, 0.78, 1.56, 3.125, 6.25, 12.5 and 25 ng/mL. Aliquot and freeze the AMH Cal H Stock immediately for multiple uses. Avoid repeated freeze thaws. Frozen aliquots at -20 °C are good for one year. l. The AMH concentration in the AMH calibrators H is traceable to the manufacturer's working calibrators. Values assigned by other methodologies may be different. Such differences, if present, may be caused by inter-method bias.
2. Wash Solution: Dilute wash concentrate 25-fold with deionized water. The wash solution is stable for one month at room temperature 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.

Probennahme:

• Serum 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 specimens³.

Testdurchführung:

Allow all specimens and reagents to reach room temperature and mix thoroughly by gentle inversion before use. Calibrators and unknowns should be assayed in duplicate.

1. Label the microtitration strips to be used.
2. Pipette 50 μL of the Calibrators (Cal A-G), controls and unknowns to the appropriate wells.
3. Add 50 μL of the AMH Assay Buffer to each well using a repeater pipette.
4. Incubate the plate, shaking at a fast speed (600-800 rpm) on an orbital microplate shaker, for 120 minutes at room temperature.
5. Aspirate and wash each strip 5 times with Wash Solution using an automatic microplate washer.
6. Add 100 μL of the Antibody-Biotin Conjugate RTU to each well using a repeater pipette.
7. Incubate the plate, shaking at a fast speed (600-800 rpm) on an orbital microplate shaker, for 60 minutes at room temperature.
8. Aspirate and wash each strip 5 times with the Wash Solution using an automatic microplate washer.
9. Add 100 μL of the Streptavidin-Enzyme Conjugate-RTU to each well using a repeater pipette.
10. Incubate the plate, shaking at a fast speed (600-800 rpm) on an orbital microplate shaker, for 30 minutes at room temperature.
11. Aspirate and wash each strip 5 times with the Wash Solution using an automatic microplate washer.
12. Add 100 μL of the TMB chromogen solution to each well using a precision pipette. Avoid exposure to direct sunlight.
13. Incubate the wells, shaking at 600-800 rpm on an orbital microplate shaker, for 10-12 min at room temperature. NOTE: Visually monitor the color development to optimize the incubation time.
14. Add 100 μL of the stopping solution to each well using a precision pipette. Read the absorbance of the solution in the wells within 20 minutes, using a microplate reader set to 450 nm. NOTE: While reading the absorbance of the microtitration well, it is necessary to program the zero calibrator (Cal A) as a ""Blank"".

Ergebnisberechnung:

NOTE: The results in this package insert were calculated by plotting the data on a log vs. log scale and using a cubic regression curve-fit. Other data reduction methods may give slightly different results.

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

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Vorsichtsmaßnahmen: 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 animal and/or 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, 20071. WARNING: Potential Chemical Hazard Some reagents in this kit contain Pro-Clean 400 and Sodium azide2 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.

Lagerung: 4 °C

Details zu AMH

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

Andere Bezeichnung: AMH (AMH Produkte)

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

Substanzklasse: Antibody

Pathways: Negative Regulation of Hormone Secretion