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Description
Mouse HBsAg 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: 1. 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: 1. 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. 2. 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. 3. 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. 4. 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. 5. 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. 6. 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 100 ng/mL). Then dilute to the following concentrations: 100 ng/mL, 50 ng/mL, 25 ng/mL, 12.5 ng/mL, 6.25 ng/mL, 3.125 ng/mL, 1.5625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each tube. Pipette 500uL of the 100ng/mL standard working solution into the first EP tube and mix thoroughly to make a 50ng/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 capture antibody against Hepatitis B surface antigen (HBsAg). 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 Hepatitis B surface antigen (HBsAg) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Hepatitis B surface antigen ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Hepatitis B virus surface antigen (HBsAg), also known as Australia antigen or Australia antigen, refers to the surface antigen of the hepatitis B virus. A positive antigen test indicates that the patient has been infected with the hepatitis B virus, which can be acute or chronic. The hepatitis B virus is an enveloped virus, and the surface proteins of its viral envelope serve as surface antigens, which are recognized by corresponding antibody proteins when infecting mice. | |||||||||||||||||||||||||||||||||
| 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 | 1.56-100 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.6 ★★★★★
Based on 1713 reviews
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Product Reviews
★★★★★ 5
Perfect grouping of toys for puppies
I have four eight week old Frenchie puppies and they definitely need some toys to play with and chew on. These are absolutely perfect. They're the right size for their little mouths and they have squeaks and little crinkles and all kinds of noises to keep them interested in the toys for quite a nice period of time. The price is excellent. And the size is perfect for them. It's a great value and if you're looking for something to occupy a puppy you can't go wrong with this
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Reviewed in the United States on April 14, 2026
★★★★★ 5
Skye's Absolute Favorite Ball! 🐾
Size: 1 Pack, Style: Ball
Hello, I am Skye! I am an Australian Mountain Doodle puppy, and my humans finally found a ball that can keep up with me! The Benebone Natural Rubber Dog Ball is officially my prized possession.
Why I give it five paws:
The Perfect Chew: It is made of natural rubber, which means it has this amazing, springy texture that feels so good on my teeth. It's super tough, but it still has a satisfying squish when I chomp down on it.
Funky Bounces: Because of the rubber and the design, it doesn't just roll in a boring straight line. When it hits the floor, it bounces all over the place in crazy directions, which makes chasing it so exciting.
Easy to Carry: The texture makes it really easy for me to grab with my paws and hold onto in my mouth, even when it gets a little slobbery during a long game of fetch.
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Reviewed in the United States on May 24, 2026
★★★★★ 5
So far this is the perfect alternative to other wobble toy feeders. Please make a bigger size!
Size: 1 Pack, Style: Pawbler
Why did you pick this product vs others?:
I bought the Pawbler because the two most popular wobble treat feeders on the market are plastic, and one of my dogs just absolutely destroys those in no time because he bites them and throws them rather then nudging them and then he has plastic cutting him so I am forced to throw away a costly treat toy/dispenser. I thought maybe a rubber one would work out better. So far my dogs love it, it works well spitting out their kibble, seems to be holding up and is easy to clean and feels like a good weight, not a super flimsy toy.
I would love to see it made in a slightly larger size, I think this current size is good for medium dogs. It holds just under a cup of small round kibble but I would like to see one that can hold more and is slightly larger in size.
I have 4 german shorthaired pointers, my test subject for this toy is mostly my 70 pounder who bites the toys instead of nudging them and throws them across the room, so it is taking a beating. I think for him a larger size would be helpful in deterring that behavior but I also think this may be too small for larger breeds.
Durability:
So far this toy looks just like when I bought it. It has no punctures or scratches and unscrews without problem. We have only had it a week or so but we use puzzle/toy feeders to feed our dogs lunch every day so it is pretty easy to tell which toys won't last long. Obviously, I can't speak to long-term durability at the moment, but I can say that most toys similar to this end up already scratched or punctured in a few days, so this is already sturdier when compared to what I have tried before.
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Reviewed in the United States on June 26, 2025
★★★★★ 4
Well made and durable, not a toy the pups seek out
Size: 1 Pack, Style: Tug
Well made. Thought our staffies would love since they live tugging with each other. Sadly not something they seek out. We love ALL the benebone chews though!
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Reviewed in the United States on April 6, 2026
★★★★★ 5
Durable fun enrichment toy
Size: 1 Pack, Style: Pawbler
I had been wanting to buy one of these kinds of toys for some time but thought that the other brands I had seen were too pricey so this was a great compromise for the price. My 5 yr old Portuguese water dog loves it. He spent 25 min playing with it the first day. The trick is to find a treat that is just big enough where it doesn’t fall out right away so it makes it challenging and fun for them. He has been chewing on it a lot for a week and it has not broken or shown signs of damage. One thing to note is that there is a small hole on the bottom of the toy so you cannot freeze liquids in it without covering the hole in some way so that would be my only complaint. Besides that it’s a great toy that keeps him busy and happy.
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Reviewed in the United States on September 1, 2025