AMH ELISA Kit

Catalog No. ABIN956178

The Human AMH ELISA Kit (ABIN956178) is a Colorimetric ELISA Kit designed to quantify Human AMH.

Quick Overview for AMH ELISA Kit (ABIN956178)

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

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Product Details AMH ELISA Kit

Purpose: This AMH ELISA kit is intended for laboratory research use only and is not for use in diagnostic or therapeutic procedures.

Analytical Method: Quantitative

Characteristics: This AMH enzyme linked immunosorbent assay applies a technique called a quantitative sandwich immunoassay. The microtiter plate provided in this kit has been pre-coated with a polyclonal antibody specific for AMH. Calibrators or samples are then added to the microtiter plate wells and AMH if present, will bind to the antibody pre-coated wells. In order to quantitatively determine the amount of AMH present in the sample, a standardized preparation of horseradish peroxidase (HRP)-conjugated polyclonal antibody, specific for AMH are added to each well to sandwich the AMH immobilized on the plate. The microtiter plate undergoes incubation, and then the wells are thoroughly washed to remove all unbound components. Next, A and B substrate solution is added to each well. The enzyme (HRP) and substrate are allowed to react over a short incubation period. Only those wells that contain AMH and enzyme- conjugated antibody will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm.

Components: Microtiter Plate: 96 wells
Calibrator 1 (0 ng/mL)
Calibrator 2 (1 ng/mL)
Calibrator 3 (2.5 ng/mL)
Calibrator 4 (5 ng/mL)
Calibrator 5 (10 ng/mL)
Calibrator 6 (25 ng/mL)
Enzyme Conjugate (1 x 6 mL)
Substrate A (1 x 6 mL)
Substrate B (1 x 6 mL)
Stop Solution (1 x 6 mL)
Wash Buffer (100X concentrate) (1 x 6 mL)
Lysis Buffer Solution (1 x 6 mL)
Note: The lysis buffer solution is used only when the sample is cell culture fluid & body fluid & tissue homogenate, If the sample is serum or blood plasma, then the lysis buffer solution is a superfluous reagent.

Material not included: 1. Microplate reader capable of measuring absorbance at 450 nm.
2. Pipettes and pipette tips.
3. 100 mL and 1 liter graduated cylinders.
4. Calibrated adjustable precision pipettes, preferably with disposable plastic tips. (A manifold multi- channel pipette is desirable for large assays.)
5. 37°C incubator.
6. Absorbent paper.
7. Distilled or de-ionized water.
8. Data analysis and graphing software. Graph paper: linear (Cartesian), log-log or semi-log, or log-logit as desired.
9. Tubes to prepare calibrator or sample dilutions.

Application Details

Plate: Pre-coated

Assay Procedure:

It is recommended that all Calibrators and Samples be added in duplicate to the Microtiter Plate.
1. Secure the desired number of coated wells in the holder then add 50 µL of Calibrators or Samples to the appropriate well in the antibody pre-coated Microtiter Plate.
2. Add 100 µL of Conjugate to each well. Mix well. Mixing well in this step is important. Cover and incubate for 1 hour at 37°C.
3. Wash the microtiter plate using one of the specified methods indicated below:
4. Manual Washing: Remove incubation mixture by aspirating contents of the plate into a sink or proper waste container. Fill in each well completely with diluted wash solution, and then aspirate contents of the plate into a sink or proper waste container. Repeat this procedure five times for a total of five washes. After washing, invert plate, and blot dry by hitting the plate onto absorbent paper or paper towels until no moisture appears. Note: Hold the sides of the plate frame firmly when washing the plate to assure that all strips remain securely in frame. Complete removal of liquid at each step is essential to good performance.
5. Automated Washing: Wash plate five times with diluted wash solution (350-400 µL/well/wash) using an auto washer. After washing, dry the plate as above. It is recommended that the washer be set for a soaking time of 10 seconds and shaking time of 5 seconds between each wash.
6. Add 50 µL substrate A and 50 µL substrate B to each well, subsequently. Cover and incubate for 15 minutes at 20-25°C. (Avoid sunlight.)
7. Add 50 µL of stop solution to each well. Mix well.
8. Read the optical density (OD) at 450 nm using a microtiter plate reader immediately.

Restrictions: For Research Use only

Handling

Storage: 4 °C

Storage Comment: All reagents provided are stored at 4°C. Refer to the expiration date on the label.

Expiry Date: The expiry date is stated on the label.

Target Details for AMH

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

Alternative Name: Anti-Mullerian Hormone

Pathways: Negative Regulation of Hormone Secretion