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Description
Mouse bACE1 ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. 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 the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL 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 the supernatant can be assayed. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend 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 freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 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 Beta-Site APP Cleaving Enzyme 1 (bACE1) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of Beta-Site APP Cleaving Enzyme 1 (bACE1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Beta-Site APP Cleaving Enzyme 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | β-Secretase 1, also known as β-site amyloid precursor protein cleaving enzyme 1, β-site APP cleaving enzyme 1 (BACE1), membrane-associated aspartic protease 2, memapsin-2, aspartic protease 2, and ASP2, is an enzyme encoded by the BACE1 gene. BACE1 is an aspartic protease that is important for the formation of myelin sheaths in peripheral nerve cells: in mice, BACE1 expression is high during the postnatal period, when myelination occurs. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.1 ★★★★★
Based on 29 reviews
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Product Reviews
★★★★★ 5
Promising Skincare Product with Clean Ingredients
I have always loved this brand and have purchased many of their skincare products in the past, so I was excited to try their Volufiline after seeing how popular it has become on TikTok. A lot of people online claim it helps plump the skin and reduce the appearance of fine lines and wrinkles without needing injections, which definitely caught my attention. For the price, I think it is a pretty reasonable product if it actually delivers results.
From what I understand, this is not a product that gives instant results and it can take at least three months or longer of consistent use before you really notice a difference. Because of that, I cannot honestly say yet whether it works or not, but I plan to continue using it and will update my review over time if I start seeing visible changes. I also purchased a separate roller ball attachment for the bottle because it makes application much easier and more precise.
Without the roller attachment, you would mostly be applying the product with your fingers, which can apparently cause your fingers themselves to become plumper from repeated contact. As always, one of the things I appreciate most about this brand is the transparency and focus on clean ingredients. Their products always feel thoughtfully formulated, and I like knowing exactly what I am putting on my skin. Overall, I am hopeful about this product and excited to see the long-term results.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 24, 2026
★★★★★ 4
Good skincare. Just okay for wrinkles and fine lines
I’m not sure it’s filling in wrinkles but it’s not hurting either. I kind of think once your a certain age there isn’t a lot of change that happens with surface level products. It’s kind of more about slowing the aging process versus reversing it. Reversing may only happen in a surgeons office or by treatment from a medical esthetician. That being said I do think this product is offering me support by slowing down the aging process and reducing new lines and wrinkles. My skin definitely has a dewy appearance after using this product. The product is lightweight and goes on like a light oil. It is a tad greasy when it’s first applied but it does absorb well. I try to put this on when I have time to let it absorb a little but I have also been in a rush and had to go straight into makeup application after using. I didn’t have any issues with the quick transition. As with most of the Ordinary line the prices are economical for skincare. I give this 4 stars only because I didn’t really see a change/improvement in my current fine lines and wrinkles. I do think overall it’s offering some benefits. The bottle is small but the amount needed per application is small. You have to shake the product out of a little restricter cap which is kind of annoying. I think a tiny dropper would make for better dispensing and less waste. As with all my facial serums and skincare products, I always rub any remaining product on the tops of my hands.
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Reviewed in the United States on June 2, 2026
★★★★★ 5
Let’s Be Honest… I Bought This Hoping for Magic
Let’s be honest-anytime a product says “plumping” or “volume,” my expectations immediately split into two categories:
absolutely nothing happens, or
I convince myself something happened because I spent money on it.
This one actually surprised me.
The texture is lightweight but nourishing, and because it’s anhydrous, it feels a little richer without being greasy or heavy. I started using a small amount consistently in areas where my skin looked a little tired or less full, and over time my skin definitely started looking smoother, healthier, and more refreshed overall.
One thing I really appreciate is that it doesn’t just sit on top of the skin making you shiny for five minutes and then disappear into disappointment. It absorbs well, layers nicely with the rest of my routine, and leaves my skin feeling soft instead of sticky.
The biggest thing I noticed is that my skin just looks more “awake” and healthy lately-like I’ve been drinking enough water, sleeping properly, and making responsible life choices…which may or may not be true.
I’ve tried a lot of skincare products that sounded impressive and did very little. This one actually earned a permanent spot in the routine.
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Reviewed in the United States on May 11, 2026
★★★★★ 5
Good product
I've used a few different volufiline’s and I like this one best. You get a tingling sensation that you don't get from other brands. Most of them are around the same price but this brand seems to have a bit of a larger bottle. I recommend purchasing this product. My smile lines are reducing significantly. I stopped putting it on my under eyes because I read somewhere it can make them worse. You also have to continue using it to keep the changes.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 24, 2026
★★★★★ 5
This makes my face feel amazing
I love how this makes my face feel! It’s still early to tell how effective it’ll be, but I saw friends yesterday that I hadn’t seen in almost a month and they commented how well rested and useful I looked! It was pretty flattering
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 31, 2026
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