Equine Anti-Müllerian hormone (AMH) ELISA Kit

Sample

Serum

Species Reactivity

Equine

Detection Method

sELISA

Intended Use

The Anti-Müllerian hormone ELISA kit provides materials for the quantitative measurement of AMH in serum, plasma and other biological fluids.

Contents of Kit

1. AMH Calibrators A / Sample Diluent
One bottle, 11 mL, labeled AMH Cal A/Sample Diluent, containing 0 ng/mL AMH in protein based buffer and Pro-Clean 400. Store unopened at 2 – 8°C until the expiration date.

2. AMH Calibrators B - F (Lyophilized)
Five vials, labeled B-F, containing concentrations of approximately 0.09 – 15.0 ng/mL AMH in protein based buffer and Pro-Clean 400. Refer to calibration card for exact concentrations. Store unopened at 2 to 8°C until the expiration date. Reconstitute calibrators B-F with 1 mL deionized water. Solubilize, Mix well and use after reconstitution. Aliquot and Freeze in plastic vials for multiple use. Alternatively, freeze in the same vial within 2 hours of reconstitution. Avoid repeated freeze thaws. The AMH concentration in the AMH calibrators 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.

3. AMH Controls I & II (Lyophilized)
Two vials, labeled Levels I and II containing low and high AMH concentrations in protein based buffer and Pro-Clean 400. Refer to calibration card for exact concentrations. Store unopened at 2 to 8°C until the expiration date. Reconstitute control Levels I and II with 1 mL deionized water. Solubilize, Mix well and use after reconstitution. Aliquot and Freeze in plastic vials for multiple use. Alternatively, freeze in the same vial within 2 hours of reconstitution. Avoid repeated freeze thaws.

4. AMH Coated Microtitration strips
One stripholder, containing 12 strips and 96 microtitration wells with AMH antibody immobilized to the inside wall of each well. Store at 2-8°C until expiration date in the resealable pouch with a desiccant to protect from moisture.

5. AMH Assay Buffer
One bottle, 12 mL, containing a protein-based (BSA)-buffer with a nonmercury preservative. Store at 2-8°C until expiration date.

6. AMH Biotin Conjugate Ready-To-Use (RTU)
One bottle, 12 mL, containing biotinylated anti-AMH antibody in protein-based buffer with a non-mercury preservative. Store at 2-8°C until expiration date.

7. AMH Streptavidin-Enzyme Conjugate-Ready-to-Use (RTU)
One amber bottle, 12 mL, containing streptavidin-HRP (horseradish peroxidase) in a protein-based buffer and a non-mercury preservative. Store undiluted at 2-8C until expiration date.

8. TMB Chromogen Solution
One bottle, 12 mL, containing a solution of tetramethylbenzidine (TMB) in buffer with hydrogen peroxide. Store at 2-8°C until expiration date.

9. Stopping Solution
One bottle, 12 mL, containing 0.2 M sulfuric acid. Store at 2 to 30°C until expiration date.

10. Wash Concentrate A
One bottle, 60 mL, containing buffered saline with a nonionic detergent. Store at 2-30C until expiration date. Dilute 25-fold with deionized water prior to use.

Storage

2-8°C (for sealed box), please do not freeze! See kit label for expiry date.

Performance Characteristics

1. All analytical characteristics are stated in ng/mL (1 ng/mL = 7.14 pmol/L)
2. Limit of Detection (LoD):
The lowest amount of AMH/MIS in a sample that can be detected with a 95% probability (n=24) is 0.023 ng/mL. The value was determined by processing five serum samples in the range of 0.03 to 0.346 ng/mL. Twelve assay runs were performed over two days with samples run in duplicate per run.
3. Limit of Quantitation (LoQ):
The estimated minimum dose achieved at 20% total imprecision is 0.06 ng/mL. The value was determined by processing eight samples in the range of 0.03 - 2.85 ng/mL over twelve runs and two days in duplicates (n=24).

General Description

Anti-Müllerian hormone is a 140 kDa glycoprotein that is produced during normal embryogenesis by the Sertoli cells of the embryonic testis, causes the involution of the Müllerian duct, and inhibits female gonadogenesis by inducing apoptosis of target gonadal cells. It belongs to the transforming growth factor-β super family. AMH causes apoptosis of specific Anti-Müllerian inhibiting substance (MIS) receptor-bearing cells, while having no effect on cells without receptors. AMH is also expressed in granulosa cells of preantral and small antral follicles in the ovary, and AMH inhibits recruitment of primordial follicles into the pool of growing follicles, and decreases responsiveness of growing follicles to FSH.