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Mouse MT(Melatonin) ELISA Kit

  • Cat.No.:E-EL-M0788

  • Reactivity: Mouse

To Purchase E-EL-M0788

Size:
  • 96T
  • 48T
  • 24T
  • 96T*5
  • 96T*10
Price: $495
Qty:

Product Details

Properties

Assay type Competitive-ELISA
Format 96T/48T
Assay time 2.0h
Detection range 7.81-500 pg/mL
Sensitivity 4.69 pg/mL
Sample type &Sample volume serum, plasma and other biological fluids; 50μL
Specificity This kit recognizes MT in samples. No significant cross-reactivity or interference between MT and analogues was observed.
Reproducibility Both intra-CV and inter-CV are < 10%.
Application This ELISA kit applies to the in vitro quantitative determination of MT concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with MT. During the reaction, MT in the sample or standard competes with a fixed amount of MT on the solid phase supporter for sites on the Biotinylated Detection Ab specific to MT. Excess conjugate and unbound sample or standard are washed away, and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of MT in tested samples can be calculated by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.
ItemSpecificationsStorage
Micro ELISA Plate(Dismountable) 96T: 8 wells ×12 strips
48T: 8 wells ×6 strips
-20℃, 6 months
Reference Standard 96T: 2 vials
48T: 1 vial
Concentrated Biotinylated Detection Ab (100×) 96T: 1 vial, 120 μL
48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×) 96T: 1 vial, 120 μL
48T: 1 vial, 60 μL
-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent 1 vial, 20 mL 2-8°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 1 vial, 10 mL 2-8℃(Protect from light)
Stop Solution 1 vial, 10 mL 2-8℃
Plate Sealer 5 pieces
Manual 1 copy
Certificate of Analysis 1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level MT were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level MT 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) 25.80 55.20 199.00 23.60 56.90 209.10
Standard deviation 1.50 2.70 10.00 1.30 3.40 9.60
CV (%) 5.81 4.89 5.03 5.51 5.98 4.59

Recovery

The recovery of MT 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=8) 89-102 94
EDTA plasma(n=8) 91-105 97
Cell culture media(n=8) 86-98 91

Linearity

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

Target Information

SynonymsN-Acetyl-5-Methoxytryptamine
Research AreaMetabolism, Neuroscience

Assay Procedures

elisa assay procedure 1

1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 45 min at 37°C

elisa assay procedure 2

2. Aspirate and wash the plate for 3 times

elisa assay procedure 3

3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times

elisa assay procedure 4

4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C

elisa assay procedure 5

5. Add 50μL Stop Solution

elisa assay procedure 6

6. Read the plate at 450nm immediately. Calculation of the results

Citations

  1. JOURNAL OF PINEAL RESEARCH (2019) IF: 15.221
    Are cortisol and melatonin involved in the immune modulation by the light environment in pike perch Sander lucioperca?

    DOI: 10.1111/jpi.12573

    Sample: Plasma
  2. JOURNAL OF PINEAL RESEARCH (2023) IF: 12.081
    Melatonin increases susceptibility to atrial fibrillation in obesity via Akt signaling impairment in response to lipid overload

    DOI: 10.1111/jpi.12851

    Sample: Serum
  3. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA (2018) IF: 9.504
    External light activates hair follicle stem cells through eyes via an ipRGC–SCN–sympathetic neural pathway

    DOI: 10.1073/pnas.1719548115

    Sample: Plasma
  4. Theranostics (2020) IF: 8.5790
    Melatonin ameliorates necrotizing enterocolitis by preventing Th17/Treg imbalance through activation of the AMPK/SIRT1 pathway

    DOI: 10.7150/thno.45862

    PMID: 32685016

    Sample: Cell culture supernatant
  5. BIOCHEMICAL PHARMACOLOGY (2022) IF: 6.1
    Melatonin regulates the immune response and improves Sjögren’s syndrome-like symptoms in NOD/Ltj Mice

    DOI: 10.1016/j.bcp.2022.115073

    Sample: salivary gl,s
  6. Cells (2022) IF: 6.6
    Activation of MT1/MT2 to Protect Testes and Leydig Cells against Cisplatin-Induced Oxidative Stress through the SIRT1/Nrf2 Signaling Pathway

    DOI: 10.3390/cells11101690

    PMID: 35626727

    Sample: testicular tissue
  7. Biology-Basel (2022) IF: 5.168
    Alterations in Gut Microbiota and Upregulations of VPAC2 and Intestinal Tight Junctions Correlate with Anti-Inflammatory Effects of Electroacupuncture in Colitis Mice with Sleep Fragmentation

    DOI: 10.3390/biology11070962

    Sample: plasma
  8. JOURNAL OF ENDOCRINOLOGICAL INVESTIGATION (2023) IF: 5.467
    A novel combination of sitagliptin and melatonin ameliorates T2D manifestations: studies on experimental diabetic models

    DOI: 10.1007/s40618-023-02014-6

    Sample: Plasma
  9. INTERNATIONAL JOURNAL OF PHARMACEUTICS (2021) IF: 5.875
    Protective effect of melatonin entrapped PLGA nanoparticles on radiation-induced lung injury through the miR-21/TGF-β1/Smad3 pathway

    DOI: 10.1016/j.ijpharm.2021.120584

    PMID: 33887395

    Sample: Serum
  10. FISH & SHELLFISH IMMUNOLOGY (2021) IF: 4.581
    Ex vivo approach supports both direct and indirect actions of melatonin on immunity in pike-perch Sander lucioperca

    DOI: 10.1016/j.fsi.2021.03.005

    PMID: 33741521

    Sample: Cell culture supernatant
  11. Frontiers in Aging Neuroscience (2019) IF: 3.633
    Melatonin and Rapamycin Attenuate Isoflurane-Induced Cognitive Impairment Through Inhibition of Neuroinflammation by Suppressing the mTOR Signaling in the Hippocampus of Aged Mice

    DOI: 10.3389/fnagi.2019.00314

    Sample: serum
  12. Scientific Reports (2020) IF: 3.998
    Seasonal simulated photoperiods influence melatonin release and immune markers of pike perch Sander lucioperca

    DOI: 10.1038/s41598-020-59568-1

    Sample: plasma
  13. CHRONOBIOLOGY INTERNATIONAL (2019) IF: 2.562
    Melatonin research in mice: a review

    DOI: 10.1080/07420528.2019.1624373

    Sample: plasma
  14. JOURNAL OF CARDIOVASCULAR PHARMACOLOGY (2020) IF: 2.598
    Activation of Paraventricular Melatonin Receptor 2 Mediates Melatonin-Conferred Cardioprotection Against Myocardial Ischemia/Reperfusion Injury

    DOI: 10.1097/FJC.0000000000000851

    Sample: plasma
  15. CHRONOBIOLOGY INTERNATIONAL (2021) IF: 2.877
    Differential response of diurnal and nocturnal mammals to prolonged altered light-dark cycle: a possible role of mood associated endocrine, inflammatory and antioxidant system

    DOI: 10.1080/07420528.2021.1937200

    PMID: 34128442

    Sample: serum,hippocampus
  16. Sleep and Biological Rhythms (2018) IF: 0.655
    Evaluation of behavior and endocrinological changes after REM sleep deprivation-induced mania-like behavior in mice

    DOI: 10.1007/s41105-018-0166-6

    Sample: serum
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Reviews/Q&A

  • Show all
  • Reviews
  • Q&A
Q

D******ASubmitted [ Feb 16 2023 ]

Asked: Does it work on hamster?

Reply

A

adminSubmitted [ Feb 16 2023 ]

Answered: Hamsters and mice are not the same species, so a mouse melatonin kit may not work with hamster samples.

Q

R************aSubmitted [ Mar 21 2022 ]

Asked: What is the dilution factor for serum?

Reply

A

adminSubmitted [ Mar 21 2022 ]

Answered: Dear Yanagisawa-san,thanks for your message. For the E-EL-M0788, we have validated the kit with normal mouse serum samples, the reference values of samples were about 15-80pg/mL,it is recommended to be undiluted. best regards, Yumi

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