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

    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    <
    >
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience

      Catalog number:E-EL-0033

      Synonyms:5HT, Enteramine, Thrombocytin, Thrombotonin, 3-(β-Aminoethyl)-5-hydroxyindole

      Size:
      • 96T
      • 24T
      Qty:
      - +
      Price: $495

      Reactivity: Universal

      Detection Range: 15.63~1000 ng/mL

      Sensitivity: 9.38 ng/mL

      Lead Time: 7~10 daysWelcome to order from local distributors.

      Add to cart Compare Bulk request Manual

      Intended use

      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 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.

      Assay type Competitive
      Format 96T
      Assay time 2.0h
      Reactivity Universal
      Detection Method Colormetric
      Detection Range 15.63—1000 ng/mL
      Sensitivity 9.38 ng/mL
      Sample Volume 50μL
      Sample Type Serum, plasma and other biological fluids

      Specificity

      This kit recognizes ST/5-HT in samples. No significant cross-reactivity or interference between ST/5-HT and analogues was observed.

      Typical data

      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.

      Concentration(ng/mL) O.D Average
      1000 0.402
      0.426
      0.414
      500 0.528
      0.532
      0.53
      250 0.73
      0.722
      0.726
      125 1.004
      1.018
      1.011
      62.5 1.355
      1.343
      1.349
      31.25 1.674
      1.654
      1.664
      15.63 1.899
      1.901
      1.9
      0 2.217
      2.221
      2.219

      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
      C V (%) 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=5) 95-107 100
      EDTA plasma (n=5) 86-99 92
      Cell culture media (n=5) 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.

          Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
      1:2 Range (%) 90-104 86-99 97-112
      Average (%) 97 92 104
      1:4 Range (%) 83-96 92-103 96-108
      Average (%) 90 98 102
      1:8 Range (%) 84-97 86-99 93-108
      Average (%) 90 92 101
      1:16 Range (%) 84-97 94-107 94-109
      Average (%) 90 99 100

      Kit components & Storage

      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.

      Item Specifications Storage
      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).

      Other supplies required

      • Microplate reader with 450 nm wavelength filter
      • High-precision transfer pipette, EP tubes and disposable pipette tips
      • Incubator capable of maintaining 37℃
      • Deionized or distilled water
      • Absorbent paper
      • Loading slot for Wash Buffer

      Assay Procedure

      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.

      Citations

      1. Publication: Bi Q R, Hou J J, Qi P, et al. Venenum Bufonis Induces Rat Neuroinflammation By Activiating NF-κB Pathway And Attenuation Of BDNF[J]. Journal of Ethnopharmacology, 2016, 186: 103-110.
        Sample Type: Tissue homogenates
      2. Publication: Özgür B G, Aksu H, Birincioğlu M, et al. Antidepressant-Like Effects Of The Xanthine Oxidase Enzyme Inhibitor Allopurinol In Rats. A Comparison With Fluoxetine[J]. Pharmacology Biochemistry and Behavior, 2015, 138: 91-95.
        Sample Type: blood
      3. Publication: Hussein A M, Abbas K M, Abulseoud O A, et al. Effects Of Ferulic Acid On Oxidative Stress, Heat Shock Protein 70, Connexin 43, And Monoamines In The Hippocampus Of Pentylenetetrazole-Kindled Rats[J]. Canadian Journal of Physiology And Pharmacology, 2017.
        Sample Type: Tissue homogenates
      4. Publication: Jiang S H, Li J, Dong F Y, et al. Increased Serotonin Signaling Contributes To The Warburg Effect In Pancreatic Tumor Cells Under Metabolic Stress And Promotes Growth Of Pancreatic Tumors In Mice[J]. Gastroenterology, 2017, 153: 277-291.
        Sample Type: Culture supernatant, Tissue homogenates and Plasma
      5. Publication: Zhang J X, Li S R, Yao S, et al. Anticonvulsant And Sedative-Hypnotic Activity Screening Of Pearl And Nacre (Mother Of Pearl)[J]. Journal of Ethnopharmacology, 2016, 181: 229-235.
        Sample Type: Tissue homogenate
      Show AllShorten
      • Show all (3)
      • Reviews (0)
      • Q&A (3)
      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.

      ... Show All Show Less
      MSDS for ELISA

      Browsing History


        People Also Bought