ELISA Kit for Melatonin (MT)

N-Acetyl-5-Methoxytryptamine

Specificity

This assay has high sensitivity and excellent specificity for detection of Melatonin (MT).
No significant cross-reactivity or interference between Melatonin (MT) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of Melatonin (MT) and the recovery rates were calculated by comparing the measured value to the expected amount of Melatonin (MT) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 86-101 96
EDTA plasma(n=5) 85-101 93
heparin plasma(n=5) 87-94 90

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Melatonin (MT) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Melatonin (MT) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Melatonin (MT) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 83-93% 80-93% 93-101% 96-103%
EDTA plasma(n=5) 79-91% 78-96% 79-94% 99-105%
heparin plasma(n=5) 81-103% 82-99% 80-101% 87-99%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1 Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
Reagent Diluent 1×300µL Stop Solution 1×6mL
TMB Substrate 1×9mL Instruction manual 1
Wash Buffer (30 × concentrate) 1×20mL

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

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Magazine Citations
Domestic Animal Endocrinology Effects of melatonin on islet neogenesis and beta cell apoptosis in streptozotocin-induced diabetic rats: an immunohistochemical study ScienceDirect: S0739724012000112
General Assembly and Scientific Symposium Biomarkers inducing changes due to microwave exposure effect on rat brain Academin: Source
Chinese Journal Comparison of the flow characteristics for the honeycomb and labyrinth seal using numerical simulation Cnki: Source
Clinics (Sao Paulo). The therapeutic effect of a pulsed electromagnetic field on the reproductive patterns of male Wistar rats exposed to a 2.45-GHz microwave field PubMed: 21876981
Journal of Radiation Research Effects of simultaneous combined exposure to CDMA and WCDMA electromagnetic fields on serum hormone levels in rats PubMed: 23239176
Experimental Gerontology Improvement of oxidative stress and immunity by melatonin: An age dependent study in golden hamster Pubmed: 23220117
Journal of Human Reproductive Sciences Melatonin and its correlation with testosterone in polycystic ovarian syndrome NCBI: PMC3963309
Indian Journal of Experimental Biology Daily variation in melatonin level, antioxidant activity and general immune response of peripheral blood mononuclear cells and lymphoid tissues of Indian goat Capra &hellip; Pubmed: 24851409
Metabolic Syndrome Serum Melatonin Level Disturbance is Related to Metabolic Syndrome and Subclinical Arterial Dysfunction in Shift Working Healthy Men Researchgate: Source
Israel Journal of Veterinary Medicine Melatonin Administration Aids Bone Healing in Diabetic Rats Ijvm: Source
Journal of Photochemistry and Photobiology B: Biology Photoperiodic induced melatonin regulates immunity and expression pattern of melatonin receptor MT1 in spleen and bone marrow mononuclear cells of male golden hamster. Pubmed: 24090924
In Vivo. Influence of?circadian?time?and lighting conditions on expression of melatonin?receptors?1?and?2?in?murinelymphocytes. Pubmed:25189896
Biomicrofluidics. Molecularly imprinted electrochemical sensing of urinary melatonin in a microfluidic system Pubmed:25584113
International Journal of Advanced Research Localization and expression of androgen receptor (AR) and melatonin membrane receptor (MT1R) in accessory sex organs of adult male golden hamster, Mesocricetus auratus: Modulation by photoperiod. Com:Source
Journal of Nanoparticle Research Titanium oxide (TiO2) nanoparticles in induction of apoptosis and inflammatory response in brain Springer:Source
International Immunopharmacology MT1 receptor expression and AA-NAT activity in lymphatic tissue following melatonin administration in male golden hamster Pubmed:25023124
Journal of Photochemistry and Photobiology B: Biology Photoperiodic regulation of melatonin membrane receptor (MT1R) expression and steroidogenesis in testis of adult golden hamster, Mesocricetus auratus Pubmed:25255424
Andrologia Melatonin prevents dexamethasone-induced testicular oxidative stress and germ cell apoptosis in golden hamster, Mesocricetus auratus Pubmed:25277062
Biological Rhythm Research Reproductive seasonality in the Indian pygmy field mouse, Mus terricolor Tandfonline:Source
Acta Histochem Developmental changes of melatonin receptor expression in the spleen of the chicken, Gallus domesticus PubMed: 26024931
Acta Histochemica Melatonin modulates monochromatic light-induced GHRH expression in the hypothalamus and GH secretion in chicks Pubmed:26948666
Veterinary Word Effect of extended photoperiod during winter on growth and onset of puberty in Murrah buffalo heifers Pubmed:27051212
Journal of Photochemistry and Photobiology B: Biology Effect of melatonin on monochromatic light-induced T-lymphocyte proliferation in the thymus of chickens Pubmed:27203566
Asian-Australas J Anim Sci. Effect of Monochromic Light-emitting Diode Light with Different Color on the Growth and ReproductivePerformances of Breeder Geese. Pubmed:26954165
Revista Brasileira de Saúde e Produção Animal Desempenho e comportamento de suínos em fase de terminação submetidos a diferentes programas de luz S1519-99402016000200272
journal of neuroendocrinology Green light inhibits GnRH-I expression by stimulating the melatonin-GnIH pathway in the chick brain. pubmed:28295740
Revista Brasileira de Medicina do Esporte EFFECTS OF DIURNAL AND NOCTURNAL STRENUOUS EXERCISE ON SERUM MELATONIN LEVELS S1517-86922016000600436
Journal of Photochemistry and Photobiology B Role of monochromatic light on daily variation of clock gene expression in the pineal gland ofchick. pubmed:27643985
Journal of Oral Science Antioxidant effects of melatonin in heart tissue after induction of experimental periodontitis inrats. pubmed:28367898
畜牧兽医学报 单色绿光对鸡胚褪黑激素合成的影响 DOI: 10.11843/j.issn.0366-6964.2017.05.019
Oxidative Medicine and Cellular Longevity Melatonin Supplementation Lowers Oxidative Stress and Regulates Adipokines in Obese Patients on a Calorie-Restricted Diet pubmed:29142618
Pakistan veterinary journal Inhibitory Effect of Melatonin on Formation of the Chicken Primordial Follicles. ISSN: 0253-8318
Acta histochemica Melatonin modulates monochromatic light-induced melatonin receptor expression in the hypothalamus of chicks pubmed:28943001
Stem Cell Reviews and Reports A Circadian Rhythm in both Complement Cascade (ComC) Activation and Sphingosine-1-Phosphate (S1P) Levels in Human Peripheral Blood Supports a … 10.1007:s12015-018-9836-7
Photochemistry and Photobiology Effect of Monochromatic Light on Circadian Rhythm of Clock Genes in Chick Pinealocytes Pubmed:29896808
Reproduction In Domestic Animals Expression of melatonin and its related synthase and membrane receptors in the oestrous corpus luteum and corpus luteum verum of sheep Pubmed:29943511
OCCUPATIONAL HYGIENE–PERFORMANCE COFACTOR IN POWER ENGINEERING. BIOCHEMICAL RESPONSE TO EXPOSURE TO EXTREMELY LOW … id:PLUMEE201601V06S01A0004
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