ELISA Kit for Melatonin (MT) Pan-species (General) Competition ELISA

N-Acetyl-5-Methoxytryptamine

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  • ELISA Kit for Melatonin (MT) Packages (Simulation)
  • ELISA Kit for Melatonin (MT) Packages (Simulation)
  • ELISA Kit for Melatonin (MT) Results demonstration
  • CEA908Ge.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

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 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

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.

ELISA Kit for Melatonin (MT)

Test principle

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Melatonin (MT) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Melatonin (MT) and unlabeled Melatonin (MT) (Standards or samples) with the pre-coated antibody specific to Melatonin (MT). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Melatonin (MT) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Melatonin (MT) in the sample.

Citations

  • Effects of melatonin on islet neogenesis and beta cell apoptosis in streptozotocin-induced diabetic rats: an immunohistochemical studyScienceDirect: S0739724012000112
  • Biomarkers inducing changes due to microwave exposure effect on rat brainAcademin: Source
  • Comparison of the flow characteristics for the honeycomb and labyrinth seal using numerical simulationCnki: Source
  • The therapeutic effect of a pulsed electromagnetic field on the reproductive patterns of male Wistar rats exposed to a 2.45-GHz microwave fieldPubMed: 21876981
  • Effects of simultaneous combined exposure to CDMA and WCDMA electromagnetic fields on serum hormone levels in ratsPubMed: 23239176
  • Improvement of oxidative stress and immunity by melatonin: An age dependent study in golden hamsterPubmed: 23220117
  • Melatonin and its correlation with testosterone in polycystic ovarian syndromeNCBI: PMC3963309
  • 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
  • Serum Melatonin Level Disturbance is Related to Metabolic Syndrome and Subclinical Arterial Dysfunction in Shift Working Healthy MenResearchgate: Source
  • Melatonin Administration Aids Bone Healing in Diabetic RatsIjvm: Source
  • 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
  • Influence of?circadian?time?and lighting conditions on expression of melatonin?receptors?1?and?2?in?murinelymphocytes.Pubmed:25189896
  • Molecularly imprinted electrochemical sensing of urinary melatonin in a microfluidic system Pubmed:25584113
  • 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
  • Titanium oxide (TiO2) nanoparticles in induction of apoptosis and inflammatory response in brainSpringer:Source
  • MT1 receptor expression and AA-NAT activity in lymphatic tissue following melatonin administration in male golden hamsterPubmed:25023124
  • Photoperiodic regulation of melatonin membrane receptor (MT1R) expression and steroidogenesis in testis of adult golden hamster, Mesocricetus auratusPubmed:25255424
  • Melatonin prevents dexamethasone-induced testicular oxidative stress and germ cell apoptosis in golden hamster, Mesocricetus auratusPubmed:25277062
  • Reproductive seasonality in the Indian pygmy field mouse, Mus terricolorTandfonline:Source
  • Developmental changes of melatonin receptor expression in the spleen of the chicken, Gallus domesticusPubMed: 26024931
  • Melatonin modulates monochromatic light-induced GHRH expression in the hypothalamus and GH secretion in chicksPubmed:26948666
  • Effect of extended photoperiod during winter on growth and onset of puberty in Murrah buffalo heifersPubmed:27051212
  • Effect of melatonin on monochromatic light-induced T-lymphocyte proliferation in the thymus of chickensPubmed:27203566
  • Effect of Monochromic Light-emitting Diode Light with Different Color on the Growth and ReproductivePerformances of Breeder Geese.Pubmed:26954165
  • Desempenho e comportamento de suínos em fase de terminação submetidos a diferentes programas de luzS1519-99402016000200272
  • Green light inhibits GnRH-I expression by stimulating the melatonin-GnIH pathway in the chick brain.pubmed:28295740
  • EFFECTS OF DIURNAL AND NOCTURNAL STRENUOUS EXERCISE ON SERUM MELATONIN LEVELSS1517-86922016000600436
  • Role of monochromatic light on daily variation of clock gene expression in the pineal gland ofchick.pubmed:27643985
  • Antioxidant effects of melatonin in heart tissue after induction of experimental periodontitis inrats.pubmed:28367898
  • 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
  • Melatonin Supplementation Lowers Oxidative Stress and Regulates Adipokines in Obese Patients on a Calorie-Restricted Dietpubmed:29142618
  • Inhibitory Effect of Melatonin on Formation of the Chicken Primordial Follicles.ISSN: 0253-8318
  • Melatonin modulates monochromatic light-induced melatonin receptor expression in the hypothalamus of chickspubmed:28943001
  • 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
  • Effect of Monochromatic Light on Circadian Rhythm of Clock Genes in Chick PinealocytesPubmed:29896808
  • Expression of melatonin and its related synthase and membrane receptors in the oestrous corpus luteum and corpus luteum verum of sheepPubmed:29943511
  • OCCUPATIONAL HYGIENE–PERFORMANCE COFACTOR IN POWER ENGINEERING. BIOCHEMICAL RESPONSE TO EXPOSURE TO EXTREMELY LOW …id:PLUMEE201601V06S01A0004
  • A Comparative Analysis of the Metabolic and Coagulative Profiles in Patients with Idiopathic Scoliosis, Congenital Scoliosis and Healthy Controls: A Case …Pubmed: 30322254
  • cBmal1 but not cClock is associated with monochromatic green light-induced circadian rhythm of melatonin in chick pinealocytesPubmed: 30533004
  • Profile of melatonin and its receptors and synthesizing enzymes in cumulus–oocyte complexes of the developing sheep antral follicle—a potential estradiol …Pubmed: 30606208
  • Effect of pinealectomy on the circadian clock of the chick retina under different monochromatic lightsPubmed: 30663441
  • Plasma levels of melatonin in dilated cardiomyopathyPubmed: 30715754
  • Elevated Serum Melatonin under Constant Darkness Enhances Neural Repair in Spinal Cord Injury through Regulation of Circadian Clock Proteins ExpressionPubmed: 30678072
  • Melatonergic agonist regulates circadian clock genes and peripheral inflammatory and neuroplasticity markers in patients with depression and anxiety
  • Mel1c Mediated Monochromatic Light-Stimulated IGF-I Synthesis through the Intracellular Gαq/PKC/ERK Signaling PathwayPubmed: 30987295
  • Role of melatonin in sleep deprivation‐induced intestinal barrier dysfunction in micePubmed: 30929267
  • Melatonin mediates monochromatic green light-induced satellite cell proliferation and muscle growth in chick embryoPubmed: 31059537
  • Effect of the melatonin nuclear receptor RORα on monochromatic light-induced T-lymphocyte proliferation in chicken thymusPubmed: 31326445
  • Effect of melatonin on monochromatic light-induced changes in clock gene circadian expression in the chick liverPubmed: 31247384
  • Melatonin maintains mitochondrial membrane potential and decreases excessive intracellular Ca2+ levels in immature human oocytesPubmed: 31472147
  • Conditional Controlled Light/Dark Cycle Influences Exercise-Induced Benefits in a Rat Model with Osteoarthritis: In Vitro and In Vivo StudyPubmed: 31684092
  • Effect of monochromatic light on the circadian clock of cultured chick retinal tissuePubmed: 32198015
  • Saliva cortisol, melatonin levels and circadian rhythm alterations in Chinese primary school children with dyslexiaPubmed: 32028434
  • Role of melatonin in intestinal mucosal injury induced by restraint stress in micePubmed: 32298156
  • Hypoxia-mediated inhibition of cholesterol synthesis leads to disruption of nocturnal sex steroidogenesis in the gonad of koi carp, Cyprinus carpioPubmed: 33034795
  • Sleep deprivation aggravated lipopolysaccharide/d-galactosamine-induced acute liver injury by suppressing melatonin productionPubmed: 32809047
  • Effects of tryptophan, a precursor for melatonin, on IVF outcomes and Doppler parameters
  • 急性睡眠剥夺对小鼠血浆褪黑激素水平和下丘脑钟基因表达的影响
  • Influence of phytomelatonin on immune cell function in male golden hamsters
  • Investigation of the effect of training on serotonin, melatonin and hematologic parameters in adolescent basketball players34090322
  • Melatonin Promotes Heterotopic Ossification Through Regulation of Endothelial-Mesenchymal Transition in Injured Achilles Tendons in Rats33644068
  • A Green and Blue Monochromatic Light Combination Therapy Reduces Oxidative Stress and Enhances B-Lymphocyte Proliferation through Promoting ¡­33828639
  • Identification of Serotonin as a Predictive Marker for Breast Cancer Patients34045888
  • Physiological crosstalk between the Mel1a and Mel1c pathways modulates melatonin-mediated, monochromatic light combination-induced bursa B-lymphocyte?¡­
  • MiR-375-3p mediates reduced pineal function in hypoxia-ischemia brain damage34280452
  • Parameters of Oxidative Stress, Vitamin D, Osteopontin, and Melatonin in Patients with Lip, Oral Cavity, and Pharyngeal Cancer34721756
  • Melatonin Nuclear Receptors Mediate Green-and-Blue-Monochromatic-Light-Combinations-Inhibited B Lymphocyte Apoptosis in the Bursa of Chickens via …Pubmed:35453433

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