Serotonin (5-hydroxytryptamine, 5-HT), a versatile monoamine neurotransmitter, is widely distributed across the central nervous system and peripheral tissues. This molecule has emerged as a crucial regulator of diverse physiological and pathological processes, influencing mood, cognition, appetite, sleep, and cardiovascular and gastrointestinal functions. It achieves these effects by interacting with a broad family of serotonin receptors and transporters, which mediate its activity in various cellular contexts.

Accumulating evidence underscores the pivotal role of serotonin in the development and progression of numerous diseases, including psychiatric disorders such as depression, anxiety, and schizophrenia, as well as neurodegenerative diseases, cardiovascular conditions, and metabolic disorders. Furthermore, dysregulation of serotonin signaling has been implicated in the modulation of other biochemical pathways, such as inflammatory responses and neuroplasticity, further emphasizing its significance in health and disease.

The RayBio^® 5-HT ELISA Kit is a competitive enzyme immunoassay developed for rapid detection and quantitation of 5-HT in serum, plasma, cell supernatant or urine samples collected from various species. The quantity of 5-HT is determined by a known 5-HT standard curve. This kit has a high sensitivity of approximately 3.35 ng/mL. Each kit provides sufficient reagents to perform up to 96 assays, including standard curve and unknown samples.

• Strip plates and additional reagents allow for use in multiple experiments
• Quantitative protein detection
• Establishes normal range
• The best products for confirmation of antibody array data

• Microplate reader capable of measuring absorbance at 450 nm
• Precision pipettes to deliver 2 μL to 1 mL volumes
• Adjustable 1-25 mL pipettes for reagent preparation
• 100 mL and 1-liter graduated cylinders
• Absorbent paper
• Distilled or deionized water
• GraphPad Prism or SigmaPlot software (or other software which can perform four-parameter logistic regression models)
• Tubes to prepare standard or sample dilutions
• Orbital shaker
• Aluminum foil
• Plastic wrap

1. Prepare all reagents, samples and standards as instructed.
2. Add 50 μL standards or samples to appropriate well. Incubate for 10 minutes at room temperature.
3. Add 50 μL anti-5-HT to appropriate well. Incubate 1 hour at room temperature. And wash 5 times.
4. Add 100 μL prepared HRP-Secondary antibody solution. Incubate 1 hour at room temperature. And wash 5 times.
5. Add 100 μL TMB One-Step Substrate Reagent to each well. Incubate 2-10 minutes at room temperature.
6. Add 50 μL Stop Solution to each well. Read at 450 nm immediately.

The entire kit may be stored at -20°C to -80°C for up to 1 year from the date of shipment. For extended storage, it is recommended to store it at -80°C. Avoid repeated freeze-thaw cycles.