Rat AMH (Anti-Mullerian Hormone) CLIA Kit (RTES00396)

SKU:
RTES00396
Product Type:
CLIA Kit
Size:
96 Assays
Sensitivity:
18.75pg/mL
Range:
31.25-2000pg/mL
ELISA Type:
Sandwich
Synonyms:
MIF, MIH, MIS
Reactivity:
Rat
Tested Sample Types:
Serum, plasma and other biological fluids
€649

Description

ELISA Kit Technical ManualMSDS

Rat 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 Rat 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 Rat 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 Rat 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 Rat AMH. The concentration of Rat AMH in the samples can be calculated by comparing the RLU of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityRat
Detection methodChemiluminescence
Detection range31.25-2000 pg/mL
Sensitivity18.75 pg/mL
Sample volume100uL
Sample typeSerum, plasma and other biological fluids
RepeatabilityCV < 15%

Specificity

This kit recognizes Rat AMH in samples. No significant cross-reactivity or interference between Rat 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
200052038
59404
5572155693
100022627
25321
2397423946
50011630
10434
1103211004
2504993
5595
52945266
1252656
2560
26082580
62.51318
1304
13111283
31.25669
679
674646
028
28
28--

Precision

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

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

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)99.33195.40990.02106.12191.421043.02
Standard deviation10.0414.23109.7910.0320.85109.00
C V (%)10.117.2811.099.4510.8910.45

Recovery

The recovery of Rat 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)94-106101
EDTA plasma (n=5)99-111104
Cell culture media (n=5)100-113106

Linearity

Samples were spiked with high concentrations of Rat 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 (%)87-10095-11388-102
Average (%)9410394
1:4Range (%)92-10890-10588-103
Average (%)999895
1:8Range (%)93-10492-10591-108
Average (%)999999
1:16Range (%)102-11486-9899-111
Average (%)10792104

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

Rat AMH (Anti-Mullerian Hormone) CLIA Kit (RTES00396) 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 100µl of standard solutions into the standard wells.
    3. Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
    4. Add 100µl 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 100µL 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 350µL 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 100µL 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 100µL 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.

Rat AMH (Anti-Mullerian Hormone) CLIA Kit (RTES00396) Protein Information

UniProt Protein Function: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.
NCBI Summary:ligand for MIS receptor II that suppresses female primary and secondary sexual development and allows male sexual differentiation to occur [RGD, Feb 2006]
UniProt Code:P49000
NCBI GenInfo Identifier:25742760
NCBI Gene ID:25378
NCBI Accession:NP_037034.1
UniProt Related Accession:P49000
Molecular Weight:58,889 Da
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  
View AllClose