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Description
Mouse LPO ELISA KitProduct Specification Usage Experimental equipment required: 1. Microplate reader (450nm) 2. High precision pipette and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled or deionized water Sample preparation and requirements: Breast milk tearsOther biological fluids: Centrifuge at 1000xg for 20 minutes, remove the supernatant, and test. Pre test preparation: 1. Remove the test kit from the refrigerator
Product Specification
| Usage |
Experimental equipment required: 1. Microplate reader (450nm) 2. High-precision pipette and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled or deionized water Sample preparation and requirements: Breast milk tearsOther 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 a lactoperoxidase (LPO) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of lactoperoxidase (LPO) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Lactoperoxidase ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Lipid peroxidases are peroxidases secreted from the mammary glands, saliva, and other mucosal glands (including the lungs, bronchi, and nose). They function as a natural first line of defense against bacteria and viruses. Lipid peroxidases are members of the heme peroxidase family. They are encoded by the LPO gene. Lipid peroxidases catalyze the oxidation of several inorganic and organic substrates with hydrogen peroxide. These substrates include bromide and iodide, thus classifying lipid peroxidases as haloperoxidases. Another important substrate is thiocyanate. The oxidation products produced by this enzyme have potent and nonspecific bactericidal and antiviral activities, including the destruction of influenza viruses. Together with its inorganic ion substrate, hydrogen peroxide, and oxidation products, it is known as the lipid peroxidase system. Therefore, when provided with adequate iodine, LPO is considered a crucial defense against invasive bacteria and viral agents, such as influenza and SARS-CoV-2. | |||||||||||||||||||||||||||||||||
| 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 | Breast milk, tears, and other biological fluids |
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4.4 ★★★★★
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Product Reviews
★★★★★ 5
So far we're loving it
Started using this about a month ago. The price for both was cheaper than just the face one at my local store so I decided to try them. I use it for me kids as well as myself and so far it's worked great. It doesn't have an overwhelming smell and doesn't leave a greasy film like some others. My kids used it and then we're outside swimming for about 2.5 hours. They got some color on their faces but we didn't reapply so I wasn't surprised. I was outside during that time too and didn't burn at all (I wasn't swimming) where I had applied it. So far I love it and will continue using.
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Reviewed in the United States on July 9, 2022
★★★★★ 5
Protect your skin before exposing it to de sun.
I can recommend this product one in mil times is excellent, my favorite
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Reviewed in the United States on August 4, 2025
★★★★★ 5
Amazinggggg
I looooove this filters, amazing feel on the skin, smells good, easy to use, good amazing for the price .
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Reviewed in the United States on August 30, 2023
★★★★★ 5
Lovely on Face
The SPF 50 for the face is moisturizing and does not settle in lines or wrinkles. It lasts longer than others I have tried. The SPF 30 is equally nice for the body.
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Reviewed in the United States on May 14, 2023
★★★★★ 5
Recommended! A Must-Have for Sun Protection
Size: 1.5 Ounce (Pack of 1)
I've been using the Neutrogena Ultra Sheer Dry Touch SPF 50 Mineral Sunscreen Stick for a while now, and I can't imagine going out without it. This convenient stick sunscreen provides excellent sun protection without leaving behind a greasy or sticky residue.
One of the things I love most about this product is the way it sits on my skin. It gives a subtle, healthy-looking glow that's far from chalky or unnatural. The "whiteness" it provides is subtle and believable, adding a touch of radiance to my complexion. I get compliments whenever I apply this on my face!
The stick format is incredibly easy to apply and reapply throughout the day. It fits perfectly in my purse and doesn't require any messy fingers. The formula absorbs quickly and leaves my skin feeling smooth and protected.
I use this sunscreen as a touch-up after applying my regular sunscreen, especially on areas like my face and neck. It provides an extra layer of protection and ensures that I'm fully covered from the sun's harmful rays.
If you're looking for a reliable and effective sunscreen that won't leave you feeling greasy or sticky, I highly recommend the Neutrogena Ultra Sheer Dry Touch SPF 50 Mineral Sunscreen Stick. It's a must-have for anyone who wants to protect their skin from the sun while maintaining a healthy, radiant glow.
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Reviewed in the United States on September 8, 2024
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