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ST/5-HT(Serotonin/5-Hydroxytryptamine) ELISA Kit

Citations(15)

  • Cat.No.:E-EL-0033

  • Reactivity: Universal
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To Purchase E-EL-0033

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  • 96T
  • 48T
  • 24T
  • 96T*5
  • 96T*10
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Product Details

Properties

Assay type Competitive-ELISA
Format 96T/48T
Assay time 2.0h
Detection range 15.63-1000 ng/mL
Sensitivity 9.38 ng/mL
Sample type &Sample volume serum, plasma and other biological fluids; 50μL
Specificity This kit recognizes ST/5-HT in samples. No significant cross-reactivity or interference between ST/5-HT 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 ST/5-HT 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 ST/5-HT. During the reaction, ST/5-HT in the sample or standard competes with a fixed amount of ST/5-HT on the solid phase supporter for sites on the Biotinylated Detection Ab specific to ST/5-HT. 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 ST/5-HT 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 ST/5-HT were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level ST/5-HT 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 (ng/mL) 51.20 141.30 362.70 54.70 132.00 351.00
Standard deviation 3.50 5.90 15.60 3.10 6.50 16.10
CV (%) 6.84 4.18 4.30 5.67 4.92 4.59

Recovery

The recovery of ST/5-HT 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) 95-107 100
EDTA plasma(n=8) 86-99 92
Cell culture media(n=8) 91-105 98

Linearity

Samples were spiked with high concentrations of ST/5-HT and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Synonyms5HT, Enteramine, Thrombocytin, Thrombotonin, 3-(β-Aminoethyl)-5-hydroxyindole

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

From now on, if you have published a paper by using any of our products since 1/1/2019, fill out the “Elabscience Publication Reward Application Form”carefully and send it to orders@elabscience.com, we will get back to you with the reward after we confirm it ASAP!

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  1. Gastroenterology (2017) IF: 18.375
    Increased Serotonin Signaling Contributes To The Warburg Effect In Pancreatic Tumor Cells Under Metabolic Stress And Promotes Growth Of Pancreatic Tumors In Mice.

    DOI: 10.1053/j.gastro.2017.03.008

    PMID: 28315323

    		 Sample: Cell culture supernatant,Tissue homogenate,Plasma
  2. European Journal of Pharmacology (2020) IF: 3.263
    Demonstration of an action pathway in mouse platelets leading to prolongation of bleeding time by fluoxetine

    DOI: 10.1016/j.ejphar.2020.173385

    PMID: 32710955

    		 Sample: Plasma,Platelet
  3. Journal of ethnopharmacology (2019) IF: 3.115
    Alkaloids from the rhizomes of Ligusticum striatum exert antimigraine effects through regulating 5-HT1B receptor and c-Jun

    DOI: 10.1016/j.jep.2019.03.026

    PMID: 30880256

    		 Sample: Tissue homogenate
  4. Journal of Ethnopharmacology (2019) IF: 3.414
    Alkaloids from the rhizomes of Ligusticum striatum exert antimigraine effects through regulating 5-HT1B receptor and c-Jun

    DOI: 10.1016/j.jep.2019.03.026

    PMID: 30880256

    		 Sample: Tissue homogenate
  5. Chinese Medicine (2020) IF: 2.96
    Hejie Zhitong prescription promotes sleep and inhibits nociceptive transmission-associated neurotransmitter activity in a rodent migraine model

    DOI: 10.1186/s13020-020-00386-y

    PMID: 33014123

    		 Sample: Whole Blood
  6. Environmental toxicology (2019) IF: 2.491
    Oxidative stress and neuromodulatory effects of deltamethrin and its combination with insect repellents in rats

    DOI: 10.1002/tox.22741

    PMID: 30884102

    		 Sample: Serum
  7. Journal of Ethnopharmacology (2016) IF: 2.981
    Anticonvulsant And Sedative-Hypnotic Activity Screening Of Pearl And Nacre (Mother Of Pearl).

    DOI: 10.1016/j.jep.2016.01.039

    PMID: 26826326

    		 Sample: Tissue homogenate
  8. Pharmacology Biochemistry and Behavior (2015) IF: 2.748
    Antidepressant-Like Effects Of The Xanthine Oxidase Enzyme Inhibitor Allopurinol In Rats. A Comparison With Fluoxetine.

    DOI: 10.1016/j.pbb.2015.09.016

    PMID: 26409178

    		 Sample: Plasma
  9. Journal of Ethnopharmacology (2016) IF: 2.981
    Venenum Bufonis Induces Rat Neuroinflammation By Activiating NF-κB Pathway And Attenuation Of BDNF.

    DOI: 10.1016/j.jep.2016.03.049

    PMID: 27013094

    		 Sample: Tissue homogenate
  10. Fisheries Science (2020) IF: 1.173
    Physiological changes in response to social isolation in male medaka fish

    DOI: 10.1007/s12562-020-01441-1

    PMID: 99999999

    		 Sample: Tissue homogenate
  11. Canadian Journal of Physiology And Pharmacology (2017) IF: 1.822
    Effects Of Ferulic Acid On Oxidative Stress, Heat Shock Protein 70, Connexin 43, And Monoamines In The Hippocampus Of Pentylenetetrazole-Kindled Rats.

    DOI: 10.1139/cjpp-2016-0219

    PMID: 28177659

    		 Sample: Tissue homogenate
  12. Journal of University of Babylon for Pure and Applied Sciences (2019)
    Measuring the Levels of Leptin and Serotonin Associated with Obesity in Adult Men in Hilla City, Iraq.

    DOI: 99999999

    PMID: 99999999

    		 Sample: Serum
  13. Turkish Journal of Science (2019)
    The Effect of Peripheral Adropin Application on Hypothalamic Pituitary Adrenal Axis in Rats

    DOI: 99999999

    PMID: 99999999

    		 Sample: Tissue homogenate
  14. Jurnal Ners (2019)
    The Affirmation–Tapping on Pain Perception and Serotonin Serum Level of Post–Caesarian Section patients

    DOI: 10.20473/jn.v14i2.16421

    PMID: 99999999

    		 Sample: Serum
  15. Research Journal of Pharmacy and Technology (2017)
    Symbiotic effect of sodium selenite and vitamin E against lithium carbonate induced brain toxicity in male rats

    DOI: 10.5958/0974-360X.2017.00293.1

    PMID: 99999999

    		 Sample: Serum
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Reviews/Q&A

  • Show all
  • Reviews
  • Q&A
Q

M***********aSubmitted [ Dec 03 2018 ]

Asked: Please there are two components in Serotonin kit : 1. Concentrated Biotinylated Detection Ab(100x) 120 μL, How can I dilute it with Biotinylated Detection Ab Diluent 14 ml ?, Or how much quality in ( ml ) can I add from Biotinylated Detection Ab Diluent to 120 μL of Concentrated Biotinylated Detection Ab(100x)? 2. Concentrated HRP Conjugate ( 100x ) 120 μL, How can I dilute it with HRP Conjugate Diluent 14 ml? Or how much quality in ( ml ) can I add from HRP Conjugate Diluent to 120 μL Concentrated HRP Conjugate ( 100x )? -- thanx--

Reply

A

adminSubmitted [ Dec 03 2018 ]

Answered: 1.Prepare the 1x Biotinylated Detection Ab working solution by add 9μL of Biotinylated Detection Ab Diluent to each microlitre of Concentrated Biotinylated Detection Ab(100x), the dilution ratio is 9:1. 2. Prepare the 1x HRP Conjugate working solution by add 9μL of HRP Conjugate Diluent to each microlitre of Concentrated HRP Conjugate( 100x ), the dilution ratio is 9:1.

Q

D*****************ySubmitted [ Aug 14 2018 ]

Asked: I Have couple of questions for the 5HT ELISA kit (E-EL-0033) you have: 1. How reliable is the serotonin antibody in terms of cross reactivity with N-acetylserotonin, melatonin, tryptamine and other biological amines? 2. What is the sensitivity as compared to the kits from other popular commercial sources like Enzo, etc? Thanks, Surajit

Reply

A

adminSubmitted [ Aug 14 2018 ]

Answered: 1. Our lab applied 5-HT coupled with BSA as the antigen to generate this kit, the antibody is mouse monoclonal with high specificity, we have also validated this kit with melatonin and tryptamine, almost no cross reactivity for the two analogues was observed. 2. The detection rang is 15.63~1000ng/mL and the sensitivity is 9.38ng/mL for our kit.

Q

e****aSubmitted [ Jul 28 2018 ]

Asked:  may i have the question whether this elisa kit is suitable for mouse ?

Reply

A

adminSubmitted [ Jul 28 2018 ]

Answered: Dear eivira, Thanks for your question. This item with cat. #E-EL-0033 is an universal ELISA kit, and it's suitable for mouse. Please feel free to contact us if any question. Thank you.

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