Mouse Developmental Biology ELISA Kits
Mouse AMH (Anti-Mullerian Hormone) CLIA Kit (MOES00079)
- SKU:
- MOES00079
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 37.5pg/mL
- Range:
- 62.5-4000pg/mL
- ELISA Type:
- Sandwich
- Synonyms:
- MIF, MIH, MIS
- Reactivity:
- Mouse
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Developmental Biology
Description
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Mouse |
| Detection method: | Chemiluminescence |
| Detection range: | 62.50-4000 pg/mL |
| Sensitivity: | 37.50 pg/mL |
| Sample volume: | 100µL |
| Sample type: | Serum, plasma and other biological fluids |
| Repeatability: | CV < 15% |
| Specificity: | This kit recognizes Mouse AMH in samples. No significant cross-reactivity or interference between Mouse AMH and analogues was observed. |
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.
| 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 |
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 |
| 4000 | 53990 64172 | 59081 | 59046 |
| 2000 | 26664 29054 | 27859 | 27824 |
| 1000 | 13597 13371 | 13484 | 13449 |
| 500 | 6594 6616 | 6605 | 6570 |
| 250 | 3274 3212 | 3243 | 3208 |
| 125 | 1636 1526 | 1581 | 1546 |
| 62.50 | 721 789 | 755 | 720 |
| 0 | 35 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 | |||||
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 204.52 | 527.86 | 1764.76 | 208.11 | 567.24 | 1626.62 |
| Standard deviation | 16.46 | 47.14 | 156.18 | 18.00 | 52.47 | 122.16 |
| C V (%) | 8.05 | 8.93 | 8.85 | 8.65 | 9.25 | 7.51 |
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) | 87-100 | 94 |
| EDTA plasma (n=5) | 99-115 | 105 |
| Cell culture media (n=5) | 94-108 | 101 |
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) | ||
| 1:2 | Range (%) | 102-114 | 98-114 | 85-99 |
| Average (%) | 108 | 105 | 91 | |
| 1:4 | Range (%) | 99-114 | 96-109 | 101-114 |
| Average (%) | 105 | 103 | 108 | |
| 1:8 | Range (%) | 99-114 | 89-104 | 100-116 |
| Average (%) | 107 | 96 | 107 | |
| 1:16 | Range (%) | 97-109 | 96-108 | 98-113 |
| Average (%) | 103 | 102 | 105 |
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 ×12 strips | -20°C, 6 months |
| Reference Standard | 2 vials | |
| Concentrated Biotinylated Detection Ab (100×) | 1 vial, 120 µL | |
| Concentrated HRP Conjugate (100×) | 1 vial, 120 µL | -20°C(shading light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 4°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | |
| HRP Conjugate Diluent | 1 vial, 14 mL | |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | |
| Substrate Reagent A | 1 vial, 5 mL | 4°C (shading light) |
| Substrate Reagent B | 1 vial, 5 mL | 4°C (shading light) |
| Plate Sealer | 5 pieces | |
| Product Description | 1 copy | |
| Certificate of Analysis | 1 copy |
- 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.
- Aliquot 100 µL of standard solutions into the standard wells.
- Add 100 µL of Sample / Standard dilution buffer into the control (zero) well.
- Add 100 µL of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids. ) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37 °C.
- 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.
- 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.
- Add 100 µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37 °C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- 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.
- Determine the RLU value of each well immediately.
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