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Description
Mouse BACE2 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 freeze-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 freeze-thaw cycles 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 1000×g for 20 minutes, and collect the supernatant for analysis. Preparation for the Assay: 1. Remove the reagent kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare a gradient standard working solution: Add 1 mL of universal diluent to the lyophilized standard. Let stand for 15 minutes to completely dissolve, then gently mix (concentration 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 seven 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 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 2 (BACE2) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of Beta-Site APP Cleaving Enzyme 2 (BACE2) 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 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| 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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4.5 ★★★★★
Based on 17 reviews
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Product Reviews
★★★★★ 5
Shark dog approved
Size: Large, Color: Broccoli, Size: Large, Color: Broccoli
My new baby shark.... I mean puppy really enjoys chewing. I can't just give her a ton of pig ears or bully sticks all day, which she would definitely love, so I'm constantly looking for well made chew toys. I'd put this right up with Kong. Very durable and nice. I also love the different textures, she seems to also love it. She can gnaw at the top and chew on the stem. She wanted it as soon as I opened it - it does smell like bacon but I haven't taken a chomp at it to be able to write a complete review!
My dog is about 23 lbs but I can see a 80 lb dog chew this, it's not small.
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Reviewed in the United States on October 8, 2025
★★★★★ 4
So cute
Size: Large, Color: Pumpkin
My French Bulldogs have really strong jaws and are aggressive chewers. These really are sturdy and last a good amount of time. They are a bit heavy and awkward for my dogs to hold onto but they do like them.
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Reviewed in the United States on April 25, 2026
★★★★★ 5
Cute toy and super tough!
Size: Small, Color: Lobster
My small dog is a serious chewer, and this lobster toy has been holding up great. Usually, he destroys toys within a day, but this one’s still going strong after a couple of weeks. The shape is perfect — easy for him to grab and chew, and the little claws keep him entertained.
The filet mignon flavor must be good because he goes right for it every time. It’s sturdy without being too hard on his teeth, and it doesn’t leave any mess or pieces behind.
If you’ve got a small but mighty chewer, this one’s definitely worth it. Cute design, lasts forever, and keeps my pup busy — total win. 🦞🐾
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Reviewed in the United States on October 26, 2025
★★★★★ 5
Excellent toy
Size: X-Large, Color: Lobster
Very durable. Dog who has destroyed every toy ever purchased loves this and it is still in great condition.
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Reviewed in the United States on April 21, 2026
★★★★★ 3
Cheese the Day with Nylabone's Power Chew Cheese Chew Toy!
Size: X-Large, Color: Cheese
As a devoted dog parent, I've always believed that our furry friends deserve the best, especially when it comes to their favorite pastime—chewing! Nylabone's Power Chew Cheese Chew Toy is a game-changer that's added an extra layer of excitement to our dog's chewing routine.
**Chew-Tastic Design:**
First things first, the cheese-inspired shape of this chew toy is a hit in our household. Our dog can't resist its quirky design, and it's become a beloved addition to his chew toy collection. It's like a fun surprise every time he reaches for it!
**Flavor Pockets for Fun:**
What truly sets this chew toy apart is its flavor pockets. We fill them with spreadable treats, turning regular chew time into an epic adventure. Our dog absolutely loves the challenge of getting every last bit of treat, and it keeps him engaged for hours.
**Dental Health Bonus:**
Beyond the fun factor, the unique textures on this toy serve a dual purpose. Not only do they provide extra sensory enjoyment for our pup, but they also help clean his teeth as he chews. It's a win-win for both entertainment and dental health.
**Built to Last:**
Now, let's talk durability. Our dog is a power chewer, and this toy has proven to be up to the challenge. Made from Nylabone's most robust material, it's stood the test of time, promoting healthy chewing habits and saving our furniture from destruction.
**Satisfaction All Around:**
This chew toy isn't just for our dog's enjoyment; it brings joy to our family too. Seeing our furry friend so engaged and happy is priceless. It satisfies his natural urge to chew while teaching him healthy chewing habits.
**A Flavorful Experience:**
Did I mention the cheese flavor? It runs throughout the toy, adding another layer of enjoyment for our pup. It's like a delicious treat that never ends!
**Perfect for Big Dogs:**
As the proud parent of an extra-large dog, we opted for the "Souper" size, designed for dogs over 50 pounds. It's the perfect size for our gentle giant.
In conclusion, Nylabone's Power Chew Cheese Chew Toy has become an integral part of our dog's life. It's more than just a chew toy; it's an adventure, a dental health solution, and hours of entertainment all rolled into one. We've embraced the mantra of "Cheese the Day" with this fantastic chew toy, and we wholeheartedly recommend it to fellow dog parents looking to add more joy and flair to their furry friend's life!
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Reviewed in the United States on October 27, 2023