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Horse 5-Hydroxytryptamine (5-HT) ELISA Kit | Precise Quantification of the Key Neurotransmitter

Product Code: YD-H-5-HT
Application: For the quantitative measurement of Horse 5-Hydroxytryptamine (5-HT, Serotonin) levels in serum, plasma, cell culture supernatants, and tissue homogenates.
Detection Range: 1.0 – 40 pg/mL
Sensitivity: < 1.0 pg/mL
Sample Volume: 10 μL
Total Assay Time: ~1.5 hours
Storage: 2-8°C, protect from light
Format: 96-well pre-coated strip plate

Product Overview: Understanding 5-Hydroxytryptamine in Equine Research

5-Hydroxytryptamine (5-HT), commonly known as serotonin, is a critical monoamine neurotransmitter and peripheral signaling molecule that regulates diverse physiological processes including mood, appetite, sleep, gastrointestinal motility, and vascular tone. In equine research, accurate quantification of Horse 5-HT is essential for studies on stress responses, behavioral disorders, gastrointestinal function, and the pathophysiology of conditions such as equine gastric ulcer syndrome (EGUS) and laminitis. Understanding serotonin dynamics in horses provides valuable insights into both welfare assessment and therapeutic interventions.

Yanda Bio’s Horse 5-Hydroxytryptamine (5-HT) ELISA Kit is meticulously developed to provide researchers and veterinarians with a sensitive and reliable tool for measuring this important biomarker. As a professional ELISA kit manufacturer with a portfolio exceeding 6,000 detectable targets and established collaborations with over 60 universities across China, we are committed to delivering high-quality, research-grade immunoassays for equine and other species.

Explore Our Horse ELISA Kit Series:


Two Optimized Kit Formats for Your Research Needs

To accommodate diverse experimental requirements and budget considerations, we offer this 5-HT ELISA kit in two specialized formats.

Format 1: Standard High-Efficiency Kit
Designed for routine, reliable quantification of 5-HT in standard research settings. This format employs a streamlined one-step incubation where antigen binds simultaneously with the solid-phase antibody and enzyme-labeled antibody. A proprietary sample diluent is included to effectively minimize interference from common matrix components such as hemoglobin and lipids, ensuring stable baseline signals and optimal binding. With extensive validation demonstrating a success rate above 99%, this format represents exceptional value for most applications.

  • Typical Pricing: Approximately $120 (retail). Bulk pricing available under $100 per kit.
  • Special Offer: Free shipping on orders of three or more ELISA kits.

Format 2: Advanced High-Sensitivity Kit
Engineered for demanding applications where sample material is precious, limited, or the target analyte is present at very low concentrations. This advanced format utilizes a biotinylated recombinant monoclonal antibody (purity >95%) paired with a high-activity Streptavidin-HRP conjugate (≥500 U/mg). This signal amplification system delivers superior sensitivity, directly circumvents matrix effects, and is designed to ensure success in critical, one-attempt experiments.

Selection Guide:

  • Choose Format 1 for cost-effective, reliable analysis in studies with sufficient sample volume
  • Choose Format 2 when working with irreplaceable samples, trace-level targets, or requiring maximum assay robustness

Assay Principle: Sandwich ELISA Technology

This kit employs a solid-phase sandwich enzyme-linked immunosorbent assay (ELISA) principle. A monoclonal antibody specific for Horse 5-Hydroxytryptamine (5-HT) is pre-coated onto the microplate. Standards and samples are pipetted into the wells, allowing 5-HT to bind to the immobilized antibody. After washing to remove unbound substances, a Horseradish Peroxidase (HRP)-conjugated detection antibody is added, forming an antibody-antigen-antibody “sandwich” complex. Following a second wash step, TMB substrate is added. The HRP enzyme catalyzes the conversion of TMB to a blue product, which turns yellow upon acidification with the stop solution. The optical density (OD) measured at 450 nm is directly proportional to the concentration of 5-HT in the sample.


Sample Collection & Preparation

Proper sample handling is critical for accurate results. Please follow these guidelines:

Sample TypeCollection & Processing
SerumUse pyrogen/endotoxin-free tubes. Avoid cell stimulation. Collect blood, centrifuge at 3000 rpm for 10 minutes to carefully separate serum from red blood cells.
PlasmaCollect using EDTA, citrate, or heparin anticoagulants. Centrifuge at 3000 rpm for 30 minutes and collect the supernatant.
Cell SupernatantCentrifuge at 3000 rpm for 10 minutes to remove particles and polymers.
Tissue HomogenateHomogenize tissue with an appropriate volume of physiological saline. Centrifuge at 3000 rpm for 10 minutes and collect the supernatant.
StorageIf samples are not tested immediately, aliquot and store at -20°C. Avoid repeated freeze-thaw cycles. Thaw at room temperature and ensure complete mixing before use.

Materials Required But Not Provided

  1. Microplate reader capable of measuring absorbance at 450 nm
  2. Precision pipettes and tips (0.5-10 μL, 2-20 μL, 20-200 μL, 200-1000 μL)
  3. 37°C incubator or water bath

Important Notes for Assay Procedure

  1. Store the kit at 2-8°C. Allow all reagents to equilibrate to room temperature for 20 minutes before use. Concentrated wash buffer may crystallize; this is normal. Dissolve crystals by warming in a water bath.
  2. Return unused strips to the resealable bag with desiccant and store properly.
  3. The S0 standard (zero concentration) serves as a negative control/blank. Note that samples are pre-diluted 5-fold during the procedure; multiply results by 5 to obtain actual concentrations.
  4. Strictly adhere to the incubation times, volumes, and sequence specified in the protocol.
  5. Mix all liquid components thoroughly before use.

Reagent Preparation

20× Wash Buffer Dilution: Dilute 1 part 20× wash buffer with 19 parts distilled water to prepare 1× working wash buffer.


Washing Method

Manual Washing: Aspirate liquid from wells, fill each well with wash buffer, allow to stand for 1 minute, then aspirate. Invert plate and tap dry on absorbent paper. Repeat 5 times.

Automated Plate Washer: Inject 350 μL of wash buffer per well, allow to soak for 1 minute, and wash 5 times.


Assay Procedure

  1. Remove the required number of strips from the aluminum foil bag after 20 minutes of room temperature equilibration. Reseal unused strips and return to 4°C.
  2. Set up standard wells and sample wells. Add 50 μL of each standard concentration to the standard wells.
  3. For sample wells: Add 10 μL of test sample followed by 40 μL of sample diluent. Leave blank wells empty.
  4. Except for blank wells, add 100 μL of HRP-conjugated detection antibody to all standard and sample wells. Seal with a plate cover and incubate at 37°C for 60 minutes.
  5. Aspirate liquid, tap dry on absorbent paper, fill each well with wash buffer, let stand for 1 minute, aspirate, and tap dry. Repeat washing 5 times (automated washer acceptable).
  6. Add 50 μL of Substrate A and 50 μL of Substrate B to each well. Incubate at 37°C in the dark for 15 minutes.
  7. Add 50 μL of stop solution to each well. Measure the OD at 450 nm within 15 minutes.

Data Analysis

Create a standard curve by plotting the OD values (y-axis) against the standard concentrations (x-axis) in Excel or suitable software. A linear regression curve (R ≥ 0.9900) provides the best fit. Calculate sample concentrations from the curve equation, multiplying by the dilution factor (5×) as indicated.


Kit Performance Specifications

ParameterPerformance
AccuracyStandard curve linear regression coefficient (R) ≥ 0.9900
SensitivityMinimum detectable concentration < 1.0 pg/mL
Detection Range1.0 – 40 pg/mL
SpecificityNo significant cross-reactivity with other monoamines or soluble structural analogs
Intra-assay CV< 15%
Inter-assay CV< 15%
Storage2-8°C, protected from light and moisture
Shelf Life6 months

Why Choose Yanda Bio?

  • Comprehensive Equine Immunoassay Portfolio: Our expanding horse ELISA series includes 5-HT for neurotransmitter research, CRP for inflammation monitoring, and IFN-γ for immune response studies—providing complete coverage for equine health and disease research.
  • Dual-Format Flexibility: Choose between standard and high-sensitivity formats to match your specific experimental needs and budget.
  • Proven Quality: Our kits are developed with rigorous validation and are trusted by researchers at more than 60 universities nationwide.
  • Reliable Support: Backed by a professional team committed to delivering high-quality products for reproducible research.
  • Custom Solutions: For specialized research needs, we offer custom ELISA kit development services. Contact us to discuss your requirements.

Discover More Horse ELISA Kits:


Special Offer

Purchase any three ELISA kits from Yanda Bio and enjoy free shipping. Combine our Horse 5-HT kit with our Horse CRP and Horse IFN-γ kits to build a comprehensive equine research panel at exceptional value.


Comparison with Premium International Brands

Yanda Bio’s ELISA kits deliver performance comparable to leading international brands at a fraction of the cost. Our efficient manufacturing process and direct-to-researcher business model eliminate unnecessary markups, allowing us to offer professional-grade immunoassays at highly competitive prices. Whether you choose our standard format for routine analysis or the high-sensitivity format for demanding applications, you receive exceptional value without compromising on data quality.


Disclaimer

Strict adherence to the protocol is required. Yanda Bio is not responsible for results obtained from deviations from the specified procedure. This product is for research use only. It is not intended for clinical diagnosis or human use. The user assumes all responsibility for any consequences arising from misuse.


Ordering & Contact Information

To order this Horse 5-Hydroxytryptamine (5-HT) ELISA Kit, or to inquire about volume discounts, format selection guidance, and custom services, please contact our sales team.

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