Human Anti-MullerianH hormone (AMH) Elisa Kit for Diagnostic Use

Human Anti-MullerianH hormone (AMH) Elisa Kit for Diagnostic Use

Intended Use
The Intended Use of this kit is to detect the concentration of the Mullerian Inhibiting Substance also called Anti-Mullerian Hormone (MIS/AMH) in the sample of Human’s serum, blood plasma, and other related tissue liquid.
 

Detection range
160 pg/ml - 5000pg/ml of MIS/AMH.
 

Test principle
The kit uses a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) to detect the level of Human Mullerian Inhibiting Substance/Anti-Mullerian hormone (MIS/AMH) in samples. In the assay, calibrators, controls and samples are incubated in microtitration wells which have been precoated with anti-AMH antibody. After incubation and washing, anti- MIS/AMH detection antibody labeled with HRP is added to each well; After additional incubation and wash, color development is carried out by adding TMB substrate, the liquid will become blue, and at the end by adding acid, the color will change to yellow.  The density of the color is positively depend on the concentration of MIS/AMH in the tested sample. By using standard curve of MIS/AMH and measuring OD450 of testing samples, the concentration of MIS/AMH in the tested sample can be correctively calculated. 
 

Materials supplied in the Kit

Note
1. This kit is not good for detecting samples containing NaN₃, because NaN₃ inhibits HRP active.
2. Specimens must be extracted and tested as soon as possible after collection.  Specimens should be kept in -20℃ but avoid repeated freeze-thaw, if not able to perform experiment after extraction.
 

Assay procedure

1.        Standards: This kit contains ready to use Standard reagents So-S6 with concentration showed below.

2.        Sample Addition and Incubation:

l   Marks wells of the Pre-coated microplate Strips for Blank (So), Standard (S1-S6) and Samples, respectively.

l   To Blank and Standard wells, add 50μl So, S1, S2, S3, S4, S5, S6, respectively.

l   To every Sample wells, add 40μl of sample diluent and 10μl of sample to be tested (5x dilution). 

l   Seal plate with sealing membrane, and incubated for 60 minutes at 37 ℃ with gently shaking.
3.Washing:
Dilute the 20ml of 20×Wash Solution (supplied) with 380ml of distilled water to make 1X wash solution. Remove the sealing membrane on the plate carefully, and drain the liquid to empty all wells by tapping plate upsidedown onto towel paper. 
To each well, add 200 ul of 1X wash solution, drain the liquid and empty all wells by tapping plate upsidedown onto towel paper. Repeat such wash 3 times.
4. Reaction: 
Add 50μl of HRP-Conjugate Reagent to each well, except the wells for Blank.  Seal plate with sealing membrane, and incubated for 60 minutes at 37 ℃ with gently shaking.
5. Washing: 
Remove the sealing membrane on the plate carefully, and drain the liquid, empty all wells by tapping plate upsidedown onto towl paper.  To each well, add 200 ul of 1X wash solution, drain the liquid and empty all wells by tapping plate upsidedown onto towl paper.  Repeat such wash 3 times.
6.  Color Development: 
To each well, add 50μl of Chromogen Solution A and 50μl of Chromogen Solution B, mix by gently shaking, then incubate for about 10 min at 37℃ (keep away from light).
7. Stop: 
When desired blue color appeared, add 50μl of Stop Solution to each well to stop the reaction (the blue color will change into yellow immediately).
8. Measurement: 
Using microplate reader, measure the optical density (OD) under 450 nm in each wells and setting the Blank well as zero.  Measurement should be carried out within 15min after adding the stop solution.
 

Rebecca Yan