ELISA Kit for 5-Hydroxytryptamine (5-HT) Pan-species (General) Competition ELISA

5HT; Serotonin; N-Methyl-Gamma

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  • ELISA Kit for 5-Hydroxytryptamine (5-HT) Packages (Simulation)
  • ELISA Kit for 5-Hydroxytryptamine (5-HT) Packages (Simulation)
  • ELISA Kit for 5-Hydroxytryptamine (5-HT) Results demonstration
  • CEA808Ge.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of 5-Hydroxytryptamine (5-HT) and the recovery rates were calculated by comparing the measured value to the expected amount of 5-Hydroxytryptamine (5-HT) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-91 81
EDTA plasma(n=5) 98-105 102
heparin plasma(n=5) 89-102 95

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level 5-Hydroxytryptamine (5-HT) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level 5-Hydroxytryptamine (5-HT) 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 5-Hydroxytryptamine (5-HT) 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) 78-97% 90-102% 78-102% 78-97%
EDTA plasma(n=5) 80-95% 78-89% 92-105% 78-98%
heparin plasma(n=5) 91-104% 78-103% 80-99% 84-95%

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.

ELISA Kit for 5-Hydroxytryptamine (5-HT)

Test principle

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to 5-Hydroxytryptamine (5-HT) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled 5-Hydroxytryptamine (5-HT) and unlabeled 5-Hydroxytryptamine (5-HT) (Standards or samples) with the pre-coated antibody specific to 5-Hydroxytryptamine (5-HT). 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 5-Hydroxytryptamine (5-HT) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of 5-Hydroxytryptamine (5-HT) in the sample.

Citations

  • Ginsenoside rich fraction of Panax ginseng C.A. Meyer improve feeding behavior following radiation-induced pica in ratsScienceDirect: S0367326X12001153
  • Sensitization induces hypersensitivity in trigeminal nervePubMed: 23106664
  • Sea buckthorn (Hippophae rhamnoides L.) oil protects against chronic stress-induced inhibitory function of natural killer cells in ratsPubMed: 26684638
  • Catha edulis chewing effects on treatment of paranoid schizophrenic patientsPubMed: 25926735
  • Electroacupuncture reCavia (Guinea pig )late hypothalamic–pituitary–adrenal axis and enhance hippocampal serotonin system in a rat model of depressionPubmed:26773866
  • Unveiling the interactions among BMPR-2, ALK-1 and 5-HTT genes in the pathophysiology of HAPEPubmed:27196063
  • Metabolic Features of Heart Failure with Different Etiology ofsp11075
  • Valproate Attenuates Nitroglycerin-Induced Trigeminovascular Activation by Preserving Mitochondrial Function in a Rat Model of Migrainepubmed:27618395
  • PI3K/AKT signaling pathway activation in a rat model of migraine.pubmed:28791398
  • The depressive-like behaviors of chronic unpredictable mild stress-treated mice, ameliorated by Tibetan medicine Zuotai: involvement in the hypothalamic …Pubmed:29379286
  • Home-Based Transcutaneous Neuromodulation Improved Constipation via Modulating Gastrointestinal Hormones and Bile AcidsPubmed:29853946
  • Changes in sexual performance and biochemical characterisation of functional neural regions: A study in serotonin transporter knockout male ratsPubmed: 31037750
  • Serotonin-Mediated Cardiac Analgesia Via Ah-Type Baroreceptor Activation Contributes to Silent Angina and Asymptomatic InfarctionPubmed: 31153963
  • TLR2-Melatonin Feedback Loop Regulates the Activation of NLRP3 Inflammasome in Murine Allergic Airway InflammationPubmed: 32117301
  • Neurotransmitter system aberrations in patients with drug addictionPubmed: 32804296
  • 重组人脑利钠肽治疗慢性肺心病急性加重期的临床效果及对5-HT 浓度的影响
  • Ganjiang granule regulates cecal microflora and serum biochemical components in a rat model of constipation-predominant irritable bowel syndrome
  • Water-Soluble Carbon Dots in Cigarette Mainstream Smoke: Their Properties and the Behavioural, Neuroendocrinological, and Neurotransmitter Changes They ¡­33762821
  • O-GlcNAc/Akt pathway regulates glucose metabolism and reduces apoptosis in liver of piglets with acute cold stress33651993
  • Investigation of the effect of training on serotonin, melatonin and hematologic parameters in adolescent basketball players34090322
  • A specific primed immune response in red palm weevil, Rhynchophorus ferrugineus, is mediated by hemocyte differentiation and phagocytosisPubmed:35245605
  • Correlation between intestinal microbiotal imbalance and 5‐HT metabolism, immune inflammation in chronic unpredictable mild stress male ratsPubmed:35535862

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