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Rat MT(Melatonin) ELISA Kit

  • Cat.No.:E-EL-R0031

  • Reactivity: Rat

To Purchase E-EL-R0031

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 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.
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) 51.50 95.00 358.50 52.50 88.90 364.10
Standard deviation 2.70 4.30 18.60 2.70 4.40 19.30
CV (%) 5.24 4.53 5.19 5.14 4.95 5.30

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) 97-110 102
EDTA plasma(n=8) 92-106 100
Cell culture media(n=8) 91-105 96

Linearity

Samples were spiked with high concentrations of Rat 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 (2020) IF: 14.528
    Measuring melatonin by immunoassay

    DOI: 10.1111/jpi.12657

    Sample: serum,plasma,biological fluids
  2. PHARMACOLOGICAL RESEARCH (2023) IF: 10.334
    Gut microbiota-derived melatonin from Puerariae Lobatae Radix-resistant starch supplementation attenuates ischemic stroke injury via a positive microbial co-occurrence pattern

    DOI: 10.1016/j.phrs.2023.106714

    Sample: brain
  3. Frontiers in Cardiovascular Medicine (2022) IF: 6.05
    Melatonin Attenuates Dasatinib-Aggravated Hypoxic Pulmonary Hypertension via Inhibiting Pulmonary Vascular Remodeling.

    DOI: 10.3389/fcvm.2022.790921

    Sample: plasma
  4. Oxidative Medicine and Cellular Longevity (2022) IF: 6.543
    Melatonin Improves the Resistance of Oxidative Stress-Induced Cellular Senescence in Osteoporotic Bone Marrow Mesenchymal Stem Cells

    DOI: 10.1155/2022/7420726

    PMID: 35087617

    Sample: serum
  5. Genes (2023) IF: 4.141
    Modification and Expression of mRNA m6A in the Lateral Habenular of Rats after Long-Term Exposure to Blue Light during the Sleep Period

    DOI: 10.3390/genes14010143

    Sample: Serum
  6. Toxics (2023) IF: 4.472
    Subchronic Low-Dose Methylmercury Exposure Accelerated Cerebral Telomere Shortening in Relevant with Declined Urinary aMT6s Level in Rats

    DOI: 10.3390/toxics11020191

    Sample: serum
  7. ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY (2020) IF: 4.872
    Dibromoacetic acid exposure is associated with abnormal melatonin rhythm in rats via inhibition of p-CREB1-AANAT signalling pathway

    DOI: 10.1016/j.ecoenv.2020.111401

    PMID: 33038730

    Sample: Serum,Tissue homogenate
  8. BASIC & CLINICAL PHARMACOLOGY & TOXICOLOGY (2017) IF: 3.176
    Prophylactic Melatonin Attenuates Isoflurane‐Induced Cognitive Impairment in Aged Rats through Hippocampal Melatonin Receptor 2 – cAMP Response Element Binding Signalling

    DOI: 10.1111/bcpt.12652

    Sample: Plasma,Tissue homogenate
  9. PLoS One (2021) IF: 3.24
    Effects of GABA/β-glucan supplements on melatonin and serotonin content extracted from natural resources

    DOI: 10.1371/journal.pone.0247890

    PMID: 33667254

    Sample: Serum
  10. PHARMACEUTICAL BIOLOGY (2022) IF: 3.503
    A comprehensive study on the relieving effect of Lilium brownii on the intestinal flora and metabolic disorder in p-chlorphenylalanine induced insomnia rats

    DOI: 10.1080/13880209.2021.2019283

    PMID: 34978949

    Sample: hypothalamus tissue
  11. PHARMACEUTICAL BIOLOGY (2020) IF: 2.971
    Lilium davidii extract alleviates p‑chlorophenylalanine‑induced insomnia in rats through modification of the hypothalamic-related neurotransmitters, melatonin and homeostasis of the hypothalamic-pituitary-adrenal axis

    DOI: 10.1080/13880209.2020.1812674

    PMID: 32924742

    Sample: brain
  12. AURIS NASUS LARYNX (2021) IF: 1.863
    Therapeutic effects of melatonin on an ovalbumin-induced allergic rhinitis model in rats

    DOI: 10.1016/j.anl.2021.04.010

    PMID: 33965284

    Sample: Serum
  13. BIOMEDICAL CHROMATOGRAPHY (2022) IF: 1.902
    Biolabel-led research pattern reveals serum profile in rats after treatment with Herba Lysimachiae: Combined analysis of metabonomics and proteomics

    DOI: 10.1002/bmc.5385

    PMID: 35445417

    Sample: serum
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