Melatonin direct Saliva (Non-Extraction) ELISA Kit

Name: Melatonin direct Saliva (Non-Extraction) ELISA Kit
SKU: DEIABL355
Rating: 5 (1 reviews)

Regulatory status: For research use only, not for use in diagnostic procedures.

COA

SDS

Datasheet

Summary

Sample

Saliva

Species Reactivity

Human

Detection Method

ELISA

Intended Use

Enzyme immunoassay for the direct, quantitative determination of melatonin in human saliva.

Contents of Kit

1. MTP Microtiter Plate Break apart strips.
MTP (12 strips of 8 wells each) coated with goat anti-rabbit antibody in solution containing bovine serum albumin, anti-rabbit IgG (goat, polyclonal). Vacuum dried.

2. CAL A and CAL B-F Standard A - F Ready to use. 0.0; 0.5; 1.5; 5.0; 15; 50; pg/mL. Contains: Melatonin and ≤ 0.1 % sodium azide (w/w).

3. CONTROL 1 & CONTROL 2
Control 1+2 Ready to use. Contains: Melatonin (low and high) and ≤ 0.1 % sodium azide (w/w). For exact concentrations see labels or QC certificate.

4. ANTISERUM Melatonin Antiserum Ready to use.
Contains: Antiserum (rabbit, polyclonal), bovine serum albumin and ≤ 0.1% ProClin (w/w).

5. BIOTIN Melatonin Biotin Ready to use.
Contains bovine serum albumin and ≤ 0.1% ProClin (w/w).

6. ENZCONJ Enzyme Conjugate Ready to use.
Contains: streptavidin conjugated to HRP, ≤ 0.02 % methylisothiazolinone (w/w) and ≤ 0.02 % bromonitrodioxane (w/w).

7. WASHBUF CONC Wash Buffer Concentrate (20x)
Phosphate buffer containing ≤ 0.2 % Tween 20 (w/w) and ≤ 0.02% ProClin (w/w).

8. TMB SUBS TMB Substrate Solution Ready to use.
Contains 3,3',5,5' Tetramethylbenzidine solution.

9. TMB STOP TMB Stop Solution Ready to use.
Contains 1 M sulfuric acid

10. FOIL Adhesive Foil

Storage

The kit is shipped at ambient temperature and should be stored at 2 - 8°C. Keep away from heat or direct sunlight. The storage and stability of specimens and prepared reagents are stated in the corresponding chapters.
The unopened reagents are stable until the expiry date indicated. The kit is stable up to 6 months after the first opening (not exceeding the expiry date) when the Microtiter plate is packed in a tightly closed bag, the bottles are closed with their screw caps and the kit is stored at indicated storage temperature.

Performance Characteristics

Limit of Blank (LoB)
The LoB study was performed with the zero calibrator (Standard A), measured in 28 replicates in one run. Limit of Blank = 0.4 pg/mL.

Limit of Quantitation (LoQ)
The LoQ study was performed with 10 saliva samples, measured in 10 replicates in one run. Limit of Quantitation = 0.85 pg/mL (CV = 20 %)

Linearity
The linearity study was performed measuring 5 different samples with different concentrations and a serial dilution up to 1:16. The assay showed a linear behavior up to a 1:16 dilution.

Precision

The intra/inter assay study was conducted with two day samples and three night samples (3.5 - 25 pg/mL) by using 1 kit lot for 20 days with two runs per day and replicate.
The intra-assay precision showed a mean CV of 15.2 % (range: 13.2 - 19.0 %).
The inter-assay precision showed a mean CV of 19.2 % (range: 16.5 - 23.8 %)
To establish between lot precision the following study design assaying five different samples was used: 3 different reagent lots / 5 days / 1 run per day per lot / 5 replicates per run.
The mean between lot variation was 13.4 % (range: 8.8 - 17.7%).

General Description

Melatonin is an intermediate product of tryptophan metabolism and is released into the blood-stream originating from the pineal gland. Melatonin production is regulated by the circadian timing system. In healthy individuals, it is produced in synchrony to the light/dark cycle, being tightly restricted to the night, provided it is dark. Light stimulus (mainly in the blue range) inhibits melatonin synthesis. The role as a physiological marker has been documented in review articles and textbooks. Melatonin concentrations play an important role in the regulation of sleep patterns. The start of melatonin production in the human body can we assessed via the dim light melatonin onset (DLMO). The DLMO allows characterization of the circadian rhythm secretion in multiple matrices (such as saliva). In apparently healthy individuals, DLMO occurs 2 - 3 hours before bedtime.

Disruption to healthy sleep patterns (exposure to bright light at night time or the participation in night shift work) can alter the DLMO and lead to a phase advance or phase delay of the melatonin concentration in the sample. Salivary melatonin is a useful biomarker in circadian dysregulation through shift work or exposure to bright light at night. Furthermore, time-resolved quantification of melatonin levels can provide information about diurnal type (morning versus evening).

Melatonin concentrations in circulation are highly variable. During the day, melatonin levels are very low. At night, melatonin concentrations rise. Illustrative examples for melatonin concentrations can be found in the scientific literature for example in the publications by Ba-Ali et al. and Wiesner et al. A large variety of methods to assess the onset of melatonin production has led to a formation of an expert group to establish recommendations for the calculation beginning of melatonin secretion. Laboratories should follow consensus guidelines on the generation of data for circadian pacemaking. Benloucif et al. have published a consensus statement for measuring melatonin in humans in different matrices (such as saliva). Several methods to obtain the DLMO in a consensus statement by Benloucif et al. For example, DLMO can be defined with a threshold calculated at 2 standard deviations above the average baseline of three or more pre-rise samples.

Clinicians should consider the effects of non-modifiable factors (e.g. age) and modifiable factors (lighting, seasonal change, physical activity) on the measurement of melatonin concentration produced in humans that are discussed in detail in peer-reviewed literature. Recommendations for sampling conditions are given in a consensus statement by Benloucif et al. Clinicians should consider the effects of comorbidities (ophthalmic diseases, spinal cord injuries, liver diseases and disorders, kidney diseases and disorders) on the measurement of melatonin concentration produced in humans that are discussed in detail in peerreviewed literature. Clinicians should consider the effects of drugs or nutrition supplements that either increase (melatonin, antidepressants, MAO inhibitors) or decrease (1-adrenergic blockers alpha-2 adrenergic agonist, benzodiazepines) on the measurement of melatonin concentration produced in humans that are discussed in detail in peer-reviewed literature. Laboratories should consider the effect of crossreactivity on the daytime melatonin concentrations that set the baseline for the quantification of melatonin onset.

Standard Curve

(Example. Do not use for calculation!)

Measuring Range: from 0.85 pg/mL (LoQ as functional sensitivity) to 38.4 pg/mL (highest concentration showing linear behavior < highest standard)

General Notes

Enzyme immunoassay for the direct, quantitative determination of melatonin in human saliva.

Citations

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Documents

pdf COA

pdf SDS

pdf Datasheet

QA & Reviews

Customer Reviews

References

Melatonin is a potential adjuvant to improve clinical outcomes in individuals with obesity and diabetes with coexistence of Covid-19

EUROPEAN JOURNAL OF PHARMACOLOGY

Authors: El-Missiry, Mohamed A.; El-Missiry, Ziad M. A.; Othman, Azza I.

Abstract

Coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is a newly discovered highly pathogenic virus that was declared pandemic in March 2020 by the World Health Organization. The virus affects the respiratory system, produces an inflammatory storm that causes lung damage and respiratory dysfunction. It infects humans of all ages. The Covid-19 takes a more severe course in individuals with chronic metabolic diseases such as obesity, diabetes mellitus, and hypertension. This category of persons exhibits weak immune activity and decreased levels of endogenous antioxidants. Melatonin is a multifunctional signaling hormone synthesized and secreted primarily by the pineal gland. It is a potent antioxidant with immunomodulatory action and has remarkable anti-inflammatory effects under a variety of circumstances. Regarding Covid-19 and metabolic syndrome, adequate information about the relationship between these two comorbidities is required for better management of these patients. Since Covid-19 infection and complications involve severe inflammation and oxidative stress in people with obesity and diabetes, we anticipated the inclusion of melatonin, as powerful antioxidant, within proposed treatment protocols. In this context, melatonin is a potential and promising agent to help overcome Covid-19 infection and boost the immune system in healthy persons and obese and diabetic patients. This review summarizes some evidence from recently published reports on the utility of melatonin as a potential adjuvant in Covid-19-infected individuals with diabetes and obesity.

Concussions in Ice Hockey - Moving Toward Objective Diagnoses and Point-of-care Treatment: A Review

CURRENT SPORTS MEDICINE REPORTS

Authors: Pender, Sara C.; Smith, Aynsley M.; Finnoff, Jonathan T.; Huston, John, III; Stuart, Michael J.

Abstract

The incidence of sport-related concussion coupled with a doubling of the participation rate in youth hockey over the past two decades provides impetus for the review of the most promising concussion treatment options. This narrative review summarizes the future treatment options for sport-related concussions in ice hockey, while acknowledging their generalizability to concussion in all sports. Symptom assessment, sign observation, as well as cognitive and balance testing, have historically been used to diagnose a concussion. These methods continue to improve, but the need for effective treatments is clear. Pharmacologic, transcranial light, and nutritional supplement treatment options for concussion warranting further investigation have been identified. Dimethyl fumarate is an immunomodulatory compound thought to trigger antioxidant gene expression. Memantine reduces apoptosis and astrogliosis by inhibiting the calcium influx into cells normally caused by glutamate's activation ofN-methyl-d-aspartate receptors. Thioredoxin-mimetic peptides and transcranial photobiomodulation temper the effects of the energy crisis by acting as free radical scavengers. In addition, seven neuroprotective nutritional supplements have been identified: berberine, creatine, curcumin, melatonin, omega-3 fatty acids, resveratrol, and vitamins. An estimated US $1.1 billion has been spent on unsuccessful traumatic brain injury clinical trials. As our ability to accurately diagnose concussion improves, dimethyl fumarate, memantine, thioredoxin-mimetic peptides, transcranial photobiomodulation, and nutritional supplements (berberine, creatine, curcumin, melatonin, omega-3 fatty acids, resveratrol, and vitamins) warrant further preclinical and clinical examination in advancing the treatment of sport-related concussions.