Description
| Save Time | Pre-coated 96 well plate | |
| Quick Start | Kit includes all necessary reagents | |
| Publication Ready | Reproducible and reliable results |
| Product Name: | Bovine Serotonin (5HT) ELISA Kit |
| Product Code: | BOEB1217 |
| Size: | 96 Assays |
| Alias: | Serotonin, 5HT, 5-hydroxytryptamine |
| Detection Method: | Competitive |
| Reactivity: | General |
| Range: | 0.78-50 ng/mL |
| Storage: | Please see kit components below for exact storage details |
| Note: | For Research Use Only |
| Component | Quantity | Storage |
| ELISA Microplate (Dismountable) | 8x12 strips | 4°C for 6 months |
| Lyophilized Standard | 2 | 4°C/ -20°C |
| Sample/Standard Dilution Buffer | 20ml | 4°C |
| Biotin-labeled Antibody (Concentrated) | 120ul | 4°C (Protection from light) |
| Antibody Dilution Buffer | 10ml | 4°C |
| HRP-Streptavidin Conjugate (SABC) | 120ul | 4°C (Protect from light) |
| SABC Dilution Buffer | 10ml | 4°C |
| TMB Substrate | 10ml | 4°C (Protection from light) |
| Stop Solution | 10ml | 4°C |
| Wash Buffer (25X) | 30ml | 4°C |
| Plate Sealer | 5 | - |
Other materials required:
Microplate reader with 450 nm wavelength filter, Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips, Incubator, Deionized or distilled water, Absorbent paper, Buffer reservoir.
*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
| Step | Procedure |
| 1. | Add 50µL of Standard, Blank, or Sample per well. The blank well is added with Sample diluent. Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming as possible. |
| 2. | Immediately add 50µL of Detection Reagent A working solution to each well. Cover with the Plate sealer. Gently tap the plate to ensure thorough mixing. Incubate for 1 hour at 37°C. Note: if Detection Reagent A appears cloudy warm to room temperature until solution is uniform. |
| 3. | Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 400µL) and let it sit in the well for 1-2 minutes. Complete removal of liquid at each step is essential. After the last wash, completely remove remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper. |
| 4. | Add 100µL of Detection Reagent B working solution to each well. Cover with the Plate sealer. Incubate for 45 minutes at 37°C. |
| 5. | Repeat the wash process for five times as conducted in step 3. |
| 6. | Add 90µL of Substrate Solution to each well. Cover with a new Plate sealer and incubate for 10-20 minutes at 37°C. Protect the plate from light. The reaction time can be shortened or extended according to the actual color change, but this should not exceed more than 30 minutes. When apparent gradient appears in standard wells, user should terminate the reaction. |
| 7. | Add 50µL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing. |
| 8. | Determine the optical density (OD value) of each well at once, using a micro-plate reader set to 450 nm. User should open the micro-plate reader in advance, preheat the instrument, and set the testing parameters. |
| 9. | After experiment, store all reagents according to the specified storage temperature respectively until their expiry. |
When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.
| Sample Type | Protocol |
| Serum | If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. |
| Plasma | Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit. |
| Urine & Cerebrospinal Fluid | Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid. |
| Cell culture supernatant | Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately. |
| Cell lysates | Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C. |
| Tissue homogenates | The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C. |
| Tissue lysates | Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C |
| Breast Milk | Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles. |
Bovine serotonin, also known as 5-HT (5-hydroxytryptamine), refers to the serotonin neurotransmitter found in bovine. Serotonin is a chemical messenger that plays a crucial role in various physiological processes in animals, including humans. It acts as a neurotransmitter in the central nervous system, where it helps regulate mood, appetite, sleep, and cognition.
Serotonin Function
In bovines, serotonin plays a vital role in various physiological functions. One of its key functions is coordinating the contractions of smooth muscles in the digestive tract. This coordination helps facilitate the movement of food and regulates the secretion of enzymes and fluids, ensuring optimal digestion. Serotonin also has implications in immune system modulation, influencing the activity of immune cells and contributing to the regulation of inflammation.
Serotonin Synthesis
Serotonin synthesis in bovines occurs in specialized cells called enterochromaffin cells, which are primarily located in the lining of the gut. These cells play a crucial role in producing serotonin. Serotonin synthesis begins with the amino acid tryptophan, which is obtained through the diet. Tryptophan serves as the precursor for serotonin production. Enzymes within the enterochromaffin cells convert tryptophan into 5-HTP and subsequently into serotonin. Once synthesized, serotonin is released into the bloodstream, allowing it to exert its effects on various tissues and organs throughout the body. The synthesis and release of serotonin are vital steps in the regulation of physiological processes.
What is the Bovine 5HT ELISA Kit?
The Bovine 5HT ELISA Kit is a laboratory tool designed for the quantitative detection of 5-hydroxytryptamine (serotonin) specifically in bovine samples. It provides a reliable and sensitive method to measure the concentration of serotonin in various biological samples, including serum, plasma, and tissue homogenates.
What are the expected benefits of using the Bovine 5HT ELISA Kit?
The Bovine 5HT ELISA Kit offers accurate and precise quantification of bovine serotonin levels, enabling reliable analysis of samples. The kit's sensitivity allows for the detection of low levels of serotonin, ensuring comprehensive data. Additionally, the simplicity and convenience of the kit streamline the process, saving time in the laboratory.
Where can I find more information about the Bovine 5HT ELISA Kit?
For more detailed information about the Bovine 5HT ELISA Kit, including technical specifications, performance characteristics, and ordering details, please refer to the product brochure or contact our customer support team. We are here to assist you with any inquiries you may have.
| Londono-Mendez, C., et al. | Seasonal metabolic and growth responses of grazing beef heifers divergent in residual feed intake | PLOS one 2026 | View Citation |
| Sepulveda Ramirez V. et al. | Using Bovine-Derived Lactic Acid Bacteria Early in Life as a Strategy to Improve Stress Resilience in Beef Calves | Master thesis 2026 | View Citation |
