Mouse AMH (Anti-Mullerian Hormone) CLIA Kit (MOES00079)

SKU:
MOES00079
€649

Description

ELISA Kit Technical ManualMSDS

Mouse AMH (Anti-Mullerian Hormone) CLIA Kit

This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Mouse AMH . Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Mouse AMH and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Mouse AMH, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Mouse AMH. The concentration of Mouse AMH in the samples can be calculated by comparing the RLU of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityMouse
Detection methodChemiluminescence
Detection range62.50-4000 pg/mL
Sensitivity37.50 pg/mL
Sample volume100uL
Sample typeSerum, plasma and other biological fluids
RepeatabilityCV < 15%

Specificity

This kit recognizes Mouse AMH in samples. No significant cross-reactivity or interference between Mouse AMH and analogues was observed.

Typical Data

As the RLU values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

Concentration
(pg/mL)
RLU Average Corrected
400053990
64172
5908159046
200026664
29054
2785927824
100013597
13371
1348413449
5006594
6616
66056570
2503274
3212
32433208
1251636
1526
15811546
62.50721
789
755720
035
35
35--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse AMH were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse AMH were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)204.52527.861764.76208.11567.241626.62
Standard deviation16.4647.14156.1818.0052.47122.16
C V (%)8.058.938.858.659.257.51

Recovery

The recovery of Mouse AMH spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type Range (%) Average Recovery (%)
Serum (n=5)87-10094
EDTA plasma (n=5)99-115105
Cell culture media (n=5)94-108101

Linearity

Samples were spiked with high concentrations of Mouse AMH and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

    Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
1:2Range (%)102-11498-11485-99
Average (%)10810591
1:4Range (%)99-11496-109101-114
Average (%)105103108
1:8Range (%)99-11489-104100-116
Average (%)10796107
1:16Range (%)97-10996-10898-113
Average (%)103102105

Kit Components & Storage

An unopened kit can be stored at 4'C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Item Specifications Storage
Micro CLIA Plate(Dismountable)8 wells X12 strips-20'C, 6 months
Reference Standard2 vials
Concentrated Biotinylated Detection Ab (100X)1 vial, 120 uL
Concentrated HRP Conjugate (100X)1 vial, 120 uL-20'C(shading light), 6 months
Reference Standard & Sample Diluent1 vial, 20 mL4'C, 6 months
Biotinylated Detection Ab Diluent1 vial, 14 mL
HRP Conjugate Diluent1 vial, 14 mL
Concentrated Wash Buffer (25X)1 vial, 30 mL
Substrate Reagent A1 vial, 5 mL4'C (shading light)
Substrate Reagent B1 vial, 5 mL4'C (shading light)
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Mouse AMH (Anti-Mullerian Hormone) CLIA Kit (MOES00079) Assay procedure

    1. Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
    2. Aliquot 100 µL of standard solutions into the standard wells.
    3. Add 100 µL of Sample / Standard dilution buffer into the control (zero) well.
    4. Add 100 µL of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids.) into test sample wells.
    5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37 °C.
    6. Aspirate the liquid from each well, do not wash. Immediately add 100 µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix.Incubate for 1 hour at 37 °C.
    7. Aspirate or decant the solution from the plate and add 350 µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
    8. Add 100 µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37 °C.
    9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
    10. Add 100 µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37 °C. Protect the plate from light.
    11. Determine the RLU value of each well immediately.

Mouse AMH (Anti-Mullerian Hormone) CLIA Kit (MOES00079) Protein Information

UniProt Protein Function:AMH: This glycoprotein, produced by the Sertoli cells of the testis, causes regression of the Muellerian duct. It is also able to inhibit the growth of tumors derived from tissues of Muellerian duct origin. Defects in AMH are the cause of persistent Muellerian duct syndrome type 1 (PMDS1). PMDS1 is a form of male pseudohermaphroditism characterized by a failure of Muellerian duct regression in otherwise normal males. Belongs to the TGF-beta family.
UniProt Protein Details:

Protein type:Secreted, signal peptide; Secreted

Cellular Component: cytoplasm; extracellular space

Molecular Function:hormone activity; receptor binding; transforming growth factor beta receptor binding

Biological Process: activation of NF-kappaB transcription factor; cell-cell signaling; Mullerian duct regression; preantral ovarian follicle growth; urogenital system development

NCBI Summary:This gene is a member of the transforming growth factor-beta superfamily of growth and differentiation factors. The encoded protein is produced in fetal Sertoli cells during male gonadal differentiation. The protein binds to the anti-Mullerian hormone receptor type 2 and causes the regression of Mullerian ducts in developing male embryos. In humans, some mutations in this gene are associated with persistent Mullerian duct syndrome (PMDS). [provided by RefSeq, Apr 2013]
UniProt Code:P27106
NCBI GenInfo Identifier:127110
NCBI Gene ID:11705
NCBI Accession:P27106.1
UniProt Related Accession:P27106
Molecular Weight:11.3 kDa
NCBI Full Name:Muellerian-inhibiting factor
NCBI Synonym Full Names:anti-Mullerian hormone
NCBI Official Symbol:Amh
NCBI Official Synonym Symbols:MIS
NCBI Protein Information:muellerian-inhibiting factor
UniProt Protein Name:Muellerian-inhibiting factor
UniProt Synonym Protein Names:Anti-Muellerian hormone; AMH; Muellerian-inhibiting substance; MIS
Protein Family:Muellerian-inhibiting factor
UniProt Gene Name:Amh
UniProt Entry Name:MIS_MOUSE
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Additional Information

Product type:
ELISA
Reactivity:
Mouse
ELISA Type:
CLIA
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