Nori Equine AMH ELISA Kit Summary

Alternative names for AMH: Anti-Müllerian hormone, Müllerian-inhibiting hormone, MIH, MIF

Alternative names for equine: Horse

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background: 

Anti-Müllerian hormone (AMH), also known as Müllerian-inhibiting hormone (MIH), is a glycoprotein hormone structurally related to inhibin and activin from the transforming growth factor beta superfamily, whose key roles are in growth differentiation and folliculogenesis. AMH is activated by SOX9 in the Sertoli cells of the male fetus.^[1] Its expression inhibits the development of the female reproductive tract, or Müllerian ducts, in the male embryo, thereby arresting the development of fallopian tubes, uterus, and upper vagina.^[2] AMH expression is critical to sex differentiation at a specific time during fetal development, and appears to be tightly regulated by nuclear receptor SF-1, transcription GATA factors, sex-reversal gene DAX1, and follicle-stimulating hormone (FSH).^[3][4] Mutations in both the AMH gene and the type II AMH receptor have been shown to cause the persistence of Müllerian derivatives in males that are otherwise normally masculinized. AMH is also a product of granulosa cells of the preantral and small antral follicles in women. As such, AMH is only present in the ovary until menopause. Production of AMH regulates folliculogenesis by inhibiting recruitment of follicles from the resting pool in order to select for the dominant follicle, after which the production of AMH diminishes.^[5] AMH can also serve as a biomarker for relative size of the ovarian reserve.^[6] In bovine, AMH can be used for selection of females in multi-ovulatory embryo transfer programs by predicting the number of antral follicles developed to ovulation.^[7] AMH can also be used as a marker for ovarian dysfunction, such as in women with polycystic ovary syndrome (PCOS).

References

1. Taguchi O, et al. (1984). Developmental Biology. 106 (2): 394–8.
2. Panidis D, et al. (2011). Medical Hypotheses. 77 (4): 649–53.
3. Shen WH, et al. (1994) Cell. 77 (5): 651–61.
4. Viger RS, et al. (1998). Development. 125 (14): 2665–75.
5. Kollmann Z, et al. (2015). Reproductive Biology and Endocrinology. 13: 21.
6. Weenen C, et al. (2004). Molecular Human Reproduction. 10 (2): 77–83.
7. Rico C, et al. (2011). Biology of Reproduction. 84 (3): 560–71.