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Description
Mouse HBeAg 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 and centrifuge at 1000 x g for 15 minutes at 2-8°C within 30 minutes of collection. Remove the supernatant for testing, or store the supernatant at -20°C or -80°C, but avoid repeated freeze-thaw cycles. Other biological fluids: Centrifuge at 1000 x g for 20 minutes. Remove the supernatant for testing. Pre-Test Preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and allow it to equilibrate to room temperature. 2. Prepare the gradient working solution of the standard: Add 1 mL of universal diluent to the lyophilized standard. Let stand for 15 minutes to completely dissolve, then gently mix (concentration 1000 ng/mL). Then dilute to the following concentrations: 1000 ng/mL, 500 ng/mL, 250 ng/mL, 125 ng/mL, 62.5 ng/mL, 31.25 ng/mL, 15.625 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 1000 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 500 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 Hepatitis B virus e Antigen (HBeAg) 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 Hepatitis B virus e Antigen (HBeAg) 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 virus e AntigenELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Hepatitis B E antigen, abbreviated as HBeAg, is a soluble protein within the core particle of the hepatitis B virus. Normally, HBeAg is embedded within HBcAg. When HBcAg breaks down, HBeAg dissolves from the hepatocyte nucleus into the serum. It appears later than HBsAg but disappears earlier than HBsAg, making it the second serological antigen marker to appear after HBsAg following HBV infection. HBeAg is detected more frequently during active hepatitis B, indicating severe liver cell damage and a high degree of infectiousness. | |||||||||||||||||||||||||||||||||
| 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 | 15.6-1000ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma and other biological fluids |
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★★★★★ 5
Happy customer
Size: 14 Slim, Color: Washed Blue
My girl absolutely loves the jeans!!!! I sized up for her just incase and I’m glad I did because they fit her perfectly!!!!!!!! I sized up for her height! Perfect length!!!! Great quality!!!! Great price!!!! Jeans look just like they are pictured online!
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Reviewed in the United States on December 19, 2025
★★★★★ 5
Cute jeans
Size: 12, Color: Washed Blue, Size: 12, Color: Washed Blue
Bought these jeans for my daughter and they fit great! The material is thick, which is nice even in warm weather. They look just like the photo and have a great length to them. Very comfortable and stretchy . She loves them!
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Reviewed in the United States on October 16, 2025
★★★★★ 5
FANTASTIC FIT
Size: 16 Plus, Color: Washed Blue
I'm a 4'7" , 65 year old woman. Due to abdominal issues I needed jeans that give that extra something on the waist. I've tried on just about every brand of jeans ranging in size from adult/junior 6 to 12. If they fit on the legs the waist was huge. If they fit on the waist the legs and butt had enough room for me and a friend. I was resigned to wearing yoga pants and leggings with big shirts. My son mentioned that he loved the Amazon basics men's dress slacks, so I figured why not. First I looked at women's and juniors, and I knew from the measurements that I was still stuck. Then I remembered that about 15 years ago I'd buy girls pants (I was a size 12 in kids at the time) Looking at the sizes I knew a 12 was way too small, then I saw the 16 plus. With hesitation I bought them. Well glory be miss Agnes, THEY FIT. Comfortable, real jeans again. Sure, I have to take them up about 6 inches, but that's fine, not a problem. I'm just so happy to have real jeans again !!!
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Reviewed in the United States on November 30, 2023
★★★★★ 5
Who doesn't love a unicorn?
Size: 2T, Color: Pink/Unicorn
Cute set but runs a bit big in my opinion for a size 2T. My grandbaby is tall for her age (22 months), but the pants were really long and loose on her, granted she's not chunky, so factor that in. Overall, it's a nice set and the material is soft (especially the pants) and seems like it would keep the little ones warm.
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Reviewed in the United States on December 31, 2025
★★★★★ 5
Cute outfit for everyday.
Size: 2T, Color: Pink/Unicorn
Exactly as pictured. Good quality, not too thin but also not too thick. True to size and looks like it will keep my baby warm when she’s inside. Good price on sale! Was a steal!
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Reviewed in the United States on December 13, 2025
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