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Description
Human VAChT 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 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 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 vesicular acetylcholine transporter (VAChT) 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 vesicular acetylcholine transporter (VAChT) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Vesicular Acetylcholine Transporter ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | The vesicular acetylcholine transporter (VAChT), also known as solute carrier family 18, member 3 (SLC18A3), is encoded by the SLC18A3 gene, located within the first intron of the choline acetyltransferase gene. It is a neurotransmitter transporter responsible for loading acetylcholine into the secretory organelles of neurons, enabling its secretion. It acts as an antiporter by transporting ACh into vesicles through the exchange of protons (H+) previously pumped into the vesicles for diffusion. ACh molecules are then carried into the vesicles by the action of exiting protons. PET imaging of this transporter may provide insights into the early diagnosis of Alzheimer's disease. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.8 ★★★★★
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Product Reviews
★★★★★ 5
So Much Better than Every Other Lip Balm
It’s a little pricey, but the best lip balm I’ve ever tried. It is lightweight, moisturizes excellently, and keeps my lips soft and comfortable. I used to use Nivea but it was heavy and greasy, and I had to apply it much more often. I’ve also tried La Mer. This is just as good but only a fraction of the price.
I first bought this when I was doing Accutane. If you’ve ever been through that experience, you know your lips are miserable for months on end, and you have to apply lip balm every fifteen minutes. I tried a dozen cheaper lip balms, but finally decided to try this. It is sooo much better. I could go over an hour without feeling uncomfortable.
Even when I’m not doing Accutane, I have very sensitive lips. During the winter I have to apply other lip balms every hour. With this one I only have to apply every four hours. If I forget to bring it with me, I can go all day without my lips feeling too bad.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 1, 2023
★★★★★ 4
Lip barrier repair not just protection
I've tried avene, okeefes, aquaphor, and this is my favorite. I use adaplene gel to treat my acne and have increased my use to daily. My skin tolerated it well but my lips became very dry (even trying to avoid adaplene gel from getting on my lips), especially on the lip line. I've been looking for a lip balm that repairs the skin, not just protects it and this is my favorite. I apply it less than the others and it works the best. I like aquaphor but I think it is more protecting instead of repair. When I was searching for a lip balm I used the ordinary barrier support serum which worked but is not as convenient/easy to apply as an actual lip balm.
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Reviewed in the United States on February 9, 2026
★★★★★ 5
Best lip balm I’ve ever used.
I love LRP’s nutritic lip balm. I don’t know if the LRP makes it anymore but I can still get it from Amazon. I’ve used LRP’s products in the past and have always been very happy with the results. I don’t have troubled skin. I’m just a guy that doesn’t want to spend large sums of money on face soap and lotion. If it were up to me I’d still be using my multiwash product to wash everything head to toe, the floor, bathroom toilet and my car. But some years ago a girl I was dating begged me to start thinking about my skin. So I took her advice, we broke up but I still heed her advice and take some consideration for skin care. It’s I like nutritic because it’s light and smooth and doesn’t have any strong feminine scents. I use this twice a day like clock work. Once in the morning before I leave the house and once when I come home after I’ve showered, ready to jump into bed. The way the balm comes out is like a woman’s lipstick. The balm is inside a tube and there is a mechanism that spins the balm up ready for application. I like to retract all the way so the edge of the cover doesn’t smoosh into the balm. It’s a little pricey for a lip balm (IMO) but I think it’s worth it. I will use this for 6-9 months. That’s twice a day everyday. Oh I don’t slather it on. It’s one swipe on the upper lip and one swipe on the lower lip.
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Reviewed in the United States on September 26, 2020
★★★★★ 3
Lip Balm
La Roche-Posay Nutritic Lip Balm is an okay product. It’s definitely not horrible, and it does what a lip balm is supposed to do. It keeps lips soft and moisturized, and it feels comfortable to wear.
That said, I’m a big fan of La Roche-Posay in general, and this one didn’t really stand out to me. I didn’t notice anything special that justified the price, and honestly, I feel like I could get similar results from a much cheaper lip balm. It works, but for the price point, I don’t think it was worth it for me.
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Reviewed in the United States on December 20, 2025
★★★★★ 5
A Moisturizing, Non-Greasy Lip Balm That Doesn't Make My Lips Itch!
Sometime in the last 3 or 4 years, I developed an allergy to lanolin, which is a common ingredient in many lip balm. I've been using an all organic lip balm without lanolin, but lately my lips have started to itch and peel again. I hated to spend $16 on one lip balm, but decided to try it out.
I got it last night and have applied it a few times since then. No itching. No greasy feeling. This is definitely a winner and worth the money. Highly recommend!
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Reviewed in the United States on February 20, 2026
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