Synonyms:5HT, Enteramine, Thrombocytin, Thrombotonin, 3-(β-Aminoethyl)-5-hydroxyindole
Detection Range: 15.63~1000 ng/mL
Sensitivity: 9.38 ng/mL
This ELISA kit applies to the in vitro quantitative determination of ST/5-HT concentrations in serum, plasma and other biological fluids.
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 from the plate, and Avidin conjugated to Horseradish Peroxidase (HRP) are added to each microplate 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 change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of ST/5-HT in the samples is then determined by comparing the OD of the samples to the standard curve.
|Detection Range||15.63—1000 ng/mL|
|Sample Type||Serum, plasma and other biological fluids|
This kit recognizes ST/5-HT in samples. No significant cross-reactivity or interference between ST/5-HT and analogues was observed.
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
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|
|C V (%)||6.84||4.18||4.30||5.67||4.92||4.59|
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 (%)|
|EDTA plasma (n=5)||86-99||92|
|Cell culture media (n=5)||91-105||98|
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.
|Serum (n=5)||EDTA plasma (n=5)||Cell culture media (n=5)|
An unopened kit can be stored at 4℃ for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
|Micro ELISA Plate(Dismountable)||8 wells ×12 strips||-20℃, 6 months|
|Reference Standard||2 vials|
|Concentrated Biotinylated Detection Ab (100×)||1 vial, 120 μL|
|Concentrated HRP Conjugate (100×)||1 vial, 120 μL||-20℃(shading light), 6 months|
|Reference Standard & Sample Diluent||1 vial, 20 mL||4℃, 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||4℃(shading light)|
|Stop Solution||1 vial, 10 mL||4℃|
|Plate Sealer||5 pieces|
|Product Description||1 copy|
|Certificate of Analysis||1 copy|
Note: All reagent bottle caps must be tightened to prevent evaporation and microbial pollution.
The volume of reagents in partial shipments is a little more than the volume marked on the label, please use accurate measuring equipment instead of directly pouring into the vial(s).
1.Add 50 μL standard or sample to each well.
2.Immediately add 50 μL Biotinylated Detection Ab to each well.
3.Incubate for 45 min at 37℃. Aspirate and wash 3 times.
4.Add 100 μL HRP Conjugate to each well. Incubate for 30 min at 37℃. Aspirate and wash 5 times.
5.Add 90 μL Substrate Reagent. Incubate for 15 min at 37℃.
6.Add 50 μL Stop Solution.
7.Read at 450 nm immediately. Calculation of results.
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--
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.
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
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.
e****aSubmitted [ Jul 28 2018 ]
Asked: may i have the question whether this elisa kit is suitable for mouse ?
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.