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Human MT(Melatonin) ELISA Kit

Cat.No.:E-EL-H2016
Reactivity: Human

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Product Details

Properties

Assay type	Competitive-ELISA
Format	96T/48T
Assay time	2.0h
Detection range	15.63-1000 pg/mL
Sensitivity	9.38 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.

Item	Specifications	Storage
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)	45.90	151.70	400.20	43.50	161.20	418.60
Standard deviation	2.50	7.90	20.80	2.30	6.90	23.00
CV (%)	5.45	5.21	5.20	5.29	4.28	5.49

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)	95-109	102
EDTA plasma(n=8)	92-106	98
Cell culture media(n=8)	89-103	96

Linearity

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

Target Information

Synonyms	N-Acetyl-5-Methoxytryptamine
Research Area	Metabolism, Neuroscience

Assay Procedures

	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
	2. Aspirate and wash the plate for 3 times
	3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	5. Add 50μL Stop Solution
	6. Read the plate at 450nm immediately. Calculation of the results

Citations

JOURNAL OF PINEAL RESEARCH (2020) IF: 14.528
Measuring melatonin by immunoassay

DOI: 10.1111/jpi.12657

Sample: serum,plasma,biological fluids
Clinical and Translational Medicine (2022) IF: 11.492
Melatonin enhances osteoblastogenesis of senescent bone marrow stromal cells through NSD2-mediated chromatin remodelling

DOI: 10.1002/ctm2.746

Sample: bone marrow plasma
GENOMICS PROTEOMICS & BIOINFORMATICS (2021) IF: 7.691
Increased Expression of Colonic Mucosal Melatonin in Patients with Irritable Bowel Syndrome Correlated with Gut Dysbiosis

DOI: 10.1016/j.gpb.2020.06.013

PMID: 33607299
Sample: Cell culture supernatant(BON-1 cells)
Frontiers in Nutrition (2022) IF: 6.59
Sleep duration of lactating mothers and its relationship with feeding pattern, milk macronutrients and related serum factors: A combined longitudinal cohort and cross-sectional study

DOI: 10.3389/fnut.2022.973291

PMID: 36110402
Sample: plasma
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: serum
Nutrients (2021) IF: 5.719
Roseburia hominis Increases Intestinal Melatonin Level by Activating p-CREB-AANAT Pathway

DOI: 10.3390/nu14010117

PMID: 35010992
Sample: BON-1 cell
Frontiers in Aging Neuroscience (2021) IF: 5.75
Peripheral Clock System Abnormalities in Patients With Parkinson’s Disease

DOI: 10.3389/fnagi.2021.736026

Sample: plasma
MITOCHONDRION (2021) IF: 4.16
Yoga improves mitochondrial health and reduces severity of autoimmune inflammatory arthritis: A randomized controlled trial

DOI: 10.1016/j.mito.2021.03.004

PMID: 33741520
Sample: serum
Aging-US (2020) IF: 4.831
Melatonin exerts immunoregulatory effects by balancing peripheral effector and regulatory T helper cells in myasthenia gravis

DOI: 10.18632/aging.103785

PMID: 33136553
Sample: Serum
International Journal of Environmental Research and Public Health (2021) IF: 3.39
A Case Control Study on Serum Levels of Potential Biomarkers in Male Breast Cancer Patients

DOI: 10.3390/ijerph18094852

PMID: 34062830
Sample: Serum
AMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY (2022) IF: 3.886
Melatonin regulates proliferation, apoptosis and invasion of trophoblasts in preeclampsia by inhibiting endoplasmic reticulum stress

DOI: 10.1111/aji.13585

PMID: 35716034
Sample: serum
WIENER KLINISCHE WOCHENSCHRIFT (2022) IF: 2.275
Peculiarities of melatonin levels in preterm infants

DOI: 10.1007/s00508-022-02109-9

Sample: urine
METABOLIC BRAIN DISEASE (2019) IF: 2.411
Evidence for an association of serum melatonin concentrations with recognition and circadian preferences in patients with schizophrenia

DOI: 10.1007/s11011-019-00395-3

PMID: 30758707
Sample: Serum
Laryngoscope Investigative Otolaryngology (2021) IF: 2.458
Relationship between physical activity, objective sleep parameters, and circadian rhythm in patients with head and neck cancer receiving chemoradiotherapy: A longitudinal study

DOI: 10.1002/lio2.664

Sample: Urine
EPILEPSY & BEHAVIOR (2020) IF: 2.508
The relationship between epileptic seizure and melatonin in children

DOI: 10.1016/j.yebeh.2020.107345

Sample: blood
CHILDS NERVOUS SYSTEM (2022) IF: 1.532
Temporal profile of serum melatonin levels in paediatric pineal tumours subjected to surgery: newer insights

DOI: 10.1007/s00381-022-05601-9

PMID: 36045301
Sample: serum
PHYCOLOGIA (2019) IF: 1.976
Exogenous melatonin confers salt stress tolerance to Chlamydomonas reinhardtii (Volvocales, Chlorophyceae) by improving redox homeostasis

DOI: 10.2216/17-141.1

Sample: Cell culture supernatant
PHYCOLOGIA (2019) IF: 1.976
Enhanced melatonin production via aralkylamine N-acetyltransferase overexpression enhances NaCl resistance in transgenic Chlamydomonas reinhardtii (Volvocales, Chlorophyceae)

DOI: 10.1080/00318884.2018.1529971

Sample: Tissue homogenate

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Reviews/Q&A

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Reviews
Q&A

a********lSubmitted [ Aug 02 2023 ]

Asked: I used this kit I want to learn the intra- and inter-assay coefficients of variation were ..........% at the range of .........pg/mL, respectively. The mean recovery of melatonin was .... %, and the sensitivity of the assay was ... pg/mL. for my study could you help me drbulbul@yahoo.com

adminSubmitted [ Aug 02 2023 ]

Answered: For E-EL-H2016, the intra- and inter-assay coefficients of variation were 5.20-5.45% at the range of 45.9-400.20pg/mL and 4.28-5.49% at the range of 43.50-418.60 pg/mL, respectively. The mean serum, EDTA plasma and cell culture media recovery of melatonin was 102%, 98%,96% and the sensitivity of the assay was 9.38 pg/mL.For details, I will reply you by email. Please pay attention to your email

adminSubmitted [ Aug 02 2023 ]

Answered: For E-EL-H2016, the intra- and inter-assay coefficients of variation were 5.20-5.45% at the range of 45.9-400.20pg/mL and 4.28-5.49% at the range of 43.50-418.60 pg/mL, respectively. The mean serum, EDTA plasma and cell culture media recovery of melatonin was 102%, 98%,96% and the sensitivity of the assay was 9.38 pg/mL. For details, I will reply you by email. Please pay attention to your email

M***** Submitted [ May 23 2023 ]

Asked: How many test can be done with one kit, or we require one kit for one person

adminSubmitted [ May 23 2023 ]

Answered: Thanks for your support.We will reply you by E-mail.

j****tSubmitted [ Oct 02 2021 ]

Asked: hello i used human melatonin elisa kit for anlaysing salivary melatonin levels, i got the result values from 10 to 999, is there any mistake i made and how to interpret my results?

adminSubmitted [ Oct 02 2021 ]

Answered: Hi Juliet, thanks for your feedback, In order to help you analysis the result, can you help to fill in the after-sales-form and send us back? I will send the and the after-sales-form to your email, pls kindly help to check it.

s***********iSubmitted [ Jan 24 2021 ]

Asked: I use horse serum for induction serum shock in my cell line culture and I wonder to know that is there any cross-reactivity in the detection of melatonin by your kit between horse serum melatonin and human cell line supernatant melatonin

adminSubmitted [ Jan 24 2021 ]

Answered: MT(melatonin) is a small molecule with almost no difference in structure among different species. We suppose that there is cross-reactivity in the detection of melatonin between horse serum melatonin and human cell line supernatant melatonin.

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