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Description
Rat 5HT4R ELISA KitProduct Specification Usage Required experimental equipment: 1. Microplate reader (450nm) 2. High precision pipettes and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37C incubator 4. Distilled or deionized water Sample preparation and requirements: Tissue homogenization: Rinse the tissue with pre chilled PBS (0. 01M, pH 7. 4) to remove residual blood (lysed red blood cells in the homogenate will affect the measurement results). Weigh and
Product Specification
| Usage | Required experimental equipment: 1. Microplate reader (450nm) 2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37°C incubator 4. Distilled or deionized water Sample preparation and requirements: Tissue homogenization: Rinse the tissue with pre-chilled PBS (0.01M, pH 7.4) to remove residual blood (lysed red blood cells in the homogenate will affect the measurement results). Weigh and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL of PBS. The specific volume can be adjusted according to experimental needs and recorded. It is recommended to add protease inhibitors to the PBS) in a glass homogenizer and grind thoroughly on ice. To further lyse tissue cells, the homogenate can be sonicated or repeatedly frozen and thawed. Finally, centrifuge the homogenate at 5000×g for 5-10 minutes, and collect the supernatant for analysis. Cell Lysis Buffer: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended in 150-200 μL of PBS per 1×10^6 cells (it is recommended to add protease inhibitors to the PBS; if the cell count is very low, reduce the PBS volume appropriately). Disrupt the cells by repeated freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. Other biological fluids: Centrifuge at 1000xg for 20 minutes, remove the supernatant, and test. Pre-test preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare the standard gradient working solution: Add 1 mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 10 ng/mL). Then dilute to the following concentrations: 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, 0.15625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 10ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves as a blank well; there is no need to pipette liquid from the penultimate tube. See the figure below for details. 3. Preparation of biotinylated detection antibody working solution: Centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute 15 minutes before use. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration with universal diluent (e.g., 10uL concentrate + 990uL universal diluent). Prepare and use immediately. 4. Prepare the enzyme conjugate working solution: 15 minutes before use, centrifuge the 100× concentrated enzyme conjugate at 1000×g for 1 minute. Dilute the 100× concentrated HRP enzyme conjugate to a 1× working concentration with universal diluent (e.g., 10 μL of concentrate + 990 μL of universal diluent). Prepare immediately. 5. Prepare the 1× wash solution: Dispense 10 mL of 20× wash solution into 190 mL of distilled water (concentrated wash solution removed from the refrigerator may crystallize; this is normal. Allow to stand at room temperature until the crystals have completely dissolved before preparing). Procedure: 1. Remove the desired strips from the aluminum foil bag after equilibration at room temperature for 10 minutes. Seal the remaining strips in a ziplock bag and return to 4°C. 2. Sample addition: Add 100 μL of sample or standard of varying concentrations to the corresponding wells. Add 100 μL of universal diluent to the blank wells. Cover with a film and incubate at 37°C for 60 minutes. (Recommendation: Dilute the sample to be tested at least 1-fold with universal diluent before adding it to the ELISA plate. This will reduce the impact of matrix effects on the test results. The sample concentration should be multiplied by the corresponding dilution factor when calculating the final sample concentration. It is recommended to run replicates for all test samples and standards.) 3. Add Biotinylated Antibody: Remove the ELISA plate and discard the liquid without washing. Add 100 μL of Biotinylated Antibody Working Solution directly to each well. Cover with a film and incubate at 37°C for 60 minutes. 4. Wash: Discard the liquid and add 300 μL of 1x Wash Solution to each well. Let stand for 1 minute, shake off the wash solution, and pat dry on absorbent paper. Repeat this process three times (a plate washer can also be used). 5. Add Enzyme Conjugate Working Solution: Add 100 μL of Enzyme Conjugate Working Solution to each well. Cover with a film and incubate at 37°C for 30 minutes. 6. Washing: Discard the liquid and wash the plate five times as in step 4. 7. Adding substrate: Add 90 μL of substrate (TMB) to each well, cover with a sealing film, and incubate at 37°C in the dark for 15 minutes. 8. Adding stop solution: Remove the ELISA plate and add 50 μL of stop solution directly to each well. Immediately measure the OD value of each well at a wavelength of 450 nm. Calculating experimental results: 1. Calculate the average OD value of the standard and sample replicates and subtract the OD value of the blank well as a correction factor. Plot the standard curve of the four-parameter logistic function on double-logarithmic graph paper, with concentration as the horizontal axis and OD value as the vertical axis. 2. If the sample OD value is higher than the upper limit of the standard curve, dilute the sample appropriately and retest. Multiply the sample concentration by the corresponding dilution factor. |
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| Theory | This kit uses a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with a 5-HT4 receptor (5HT4R) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of 5-HT4 receptor (5HT4R) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat 5-HT4 Receptor ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Serotonin receptor 4 (5HT4) is a protein encoded by the HTR4 gene. This gene is a member of the serotonin receptor family and a G protein-coupled receptor that stimulates cAMP production in response to serotonin (5-hydroxytryptamine). The gene product is a glycosylated transmembrane protein that regulates the release of various neurotransmitters in the peripheral and central nervous systems. Multiple transcript variants have been described, encoding proteins with varying C-terminal sequences. | |||||||||||||||||||||||||||||||||
| General Notes | 1. Strictly adhere to the specified incubation time and temperature to ensure accurate results. All reagents must be at room temperature (20-25°C) before use. Refrigerate reagents immediately after use. 2. Improper plate washing may result in inaccurate results. Ensure that all liquid in the wells is aspirated thoroughly before adding substrate. Do not allow the wells to dry out during incubation. 3. Remove any residual liquid and fingerprints from the bottom of the plate, as this will affect the OD value. 4. The substrate developer solution should be colorless or very light in color. Do not use substrate solution that has turned blue. 5. Avoid cross-contamination of reagents and specimens to prevent erroneous results. 6. Avoid direct exposure to strong light during storage and incubation. 7. Do not expose any reagents to bleaching solvents or the strong fumes emitted by bleaching solvents. Any bleaching agent will destroy the biological activity of the reagents in the kit. 8. Do not use expired products, and do not mix components with different product numbers and batches. 9. Recombinant proteins from sources other than the kit may not be compatible with the antibodies in this kit and will not be recognized. 10. If there is a possibility of disease transmission, all samples should be managed properly and samples and testing devices should be handled according to prescribed procedures. |
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| Storage Temp. | If the unopened kit is stored at 4°C, the shelf life is 6 months. | |||||||||||||||||||||||||||||||||
| Test Range | 0.156-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.8 ★★★★★
Based on 22 reviews
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Product Reviews
★★★★★ 5
PACKS A PUNCH!!👊👊👊
Size: 21.6Inch Light Bar, Size: 21.6Inch Light Bar
Solid quality. Installed for the past month now. Been through heavy rain,freezing rain and as low as 20°. No signs of condensation as of yet. Hardware mounts and sets secure, not worried about anything coming apart. Make sure to order your wiring harness.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 5, 2024
★★★★★ 5
Very bright !
Size: 2Pcs 7.6Inch Light Bar, Size: 2Pcs 7.6Inch Light Bar
I haven't had a chance to use them on the river yet. I just finished installing them.
They were easy to install and they are very bright !
My home is at the end of a cul-de-sac and with my boat parked in the garage I can light up the house on the other side of the street.
I'll need to adjust the lights when I'm on the river. The mounting hardware supplied seems like it will be an easy task.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 9, 2023
★★★★★ 5
Straightforward setup Excellent Warm sound
Style: AVR-S770H
Setup was surprisingly easy. No manual included, but a quick link to a PDF. Connection to PC, Nintendo Switch, Cable Box done in minutes. Setup Menu worked great. Bluetooth pairing to phone easy. Setup as 5.1 (waiting on 2 more speakers for 7.1 connection). Small glitch in that the speakers didn't respond or even test tone right away. Went in and out of menu a few times, and then they worked (did this once with a 2-speaker quicker setup and then did the same glitch when adding the additional speakers to the profile with the full 5.1 setup). Works with ARC well (Samsung TV that came with silly external box) and reciever connection defaults to a passthrough when not on, which is nice. Video is great and compressed audio cleanup works well. Didn't do Mic leveling yet, but the manual speaker tuning worked well at leveling things out for 5.1. I was expected a lot harder set-up and was pleasantly surprised. It's a quality product. Some folks may not like the direct DB volume adjustment but I find it handy, especially when working a little later at night, I can dial in a very exacting low volume so as not to disturb the rest of the house. Very happy with purchase.
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Reviewed in the United States on May 28, 2024
★★★★★ 5
Great product
Style: AVR-S770H
I have owned several $1000.00 plus theater systems of 7.1(2) channels and I have never heard my speakers put out the sound like the Denon does. I would never change systems again unless Denon stops producing this product
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Reviewed in the United States on May 13, 2026
★★★★★ 5
AVR-S770H - My first AVR and my first Denon - works great with Directv and Fire Stick
Style: AVR-S770H
This review is for the Denon AVR-S770H. I was apprehensive about buying an AVR for my main TV. I've always used a soundbar and was happy with the results. I assumed that adding an AVR would make things finicky and complicated. I'm happy to say, that is not true. This Denon has been fabulous.
My requirements for a home theater receiver were/are the following:
1) Ease of use and NO additional remotes to our current set-up (family needs, I'm not a bachelor!)
2) I need the ability to connect 2 zones: the TV room and I have a pair of outdoor speakers on the patio
First, as to the remote situation, I use Directv and I use an Amazon Fire Stick 4k. My old set-up was super simple. I picked up the Directv remote and hit Power. That turned on the TV, the Directv box, and the soundbar (since it was connected to the TV ARC port). Simple. If I wanted to change over to the Fire Stick, I simply used the same Directv remote to change the input and then used the Fire Stick remote from there.
I'm happy to say the Denon has made things even simpler. Not only does the Directv remote turn on/off the TV, Direct box, and the Denon receiver but now I don't even have to bother switching inputs. If I pick up the Fire Stick remote and hit the Home button, the Denon senses that and immediately switches over. Likewise, if I pick up the Directv remote and hit a button, the Denon will switch back over to Directv. So that is an actual improvement.
By the way, in order to get the Directv remote to turn off the Denon, don't worry about trying to program the remote via the Directv menu (I tried all the codes and failed). Instead, go to the Denon setting menu. Choose Setup> Video> HDMI Setup. Change the "Power Off Control" to "Video" or "All". Now the Denon will turn on/off when the TV does (using the Directv remote).
My second concern was operating a second set of speakers via Zone 2. That also works perfectly. Now when I step outside to sit on the patio, I simply hit the Zone 2 button on the front of the Denon. I can use my phone to play music via the Denon's built-in Bluetooth.
Some users need to be aware: in order to operate Zone 2, the receiver needs to be switched from 7.2 channel to 5.1 + Zone 2. That's perfectly acceptable for me since my TV has only front L & R speakers and a subwoofer. So I'm not even using the full capabilities of the 5.1. If you need 7.2 and a second zone, you need to find a different AVR.
In sum, I bought this Denon with some trepidation and thought maybe I'd have to return it. Instead, it does everything I want and then some. Of course, it sounds great too. I'm so glad I switched from the soundbar.
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Reviewed in the United States on May 4, 2025
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