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Description
Mouse IL12B 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 200 pg/mL). Then dilute to the following concentrations: 200 pg/mL, 100 pg/mL, 50 pg/mL, 25 pg/mL, 12.5 pg/mL, 6.25 pg/mL, 3.125 pg/mL, and 0 pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 200pg/mL standard working solution into the first EP tube and mix thoroughly to make a 100pg/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 utilizes 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 Interleukin 12B (IL12B) 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 Interleukin 12B (IL12B) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Interleukin 12B ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | The beta subunit of interleukin-12 (also known as IL-12B, natural killer cell-stimulating factor, cytotoxic lymphocyte maturation factor p40, or interleukin-12 subunit p40) is a protein encoded by the IL12B gene. IL-12B is a common subunit of interleukin-12 and interleukin-23. This gene encodes a subunit of interleukin-12, a cytokine that acts on T cells and natural killer cells and exhibits a wide range of biological activities. Interleukin-12 is a disulfide-linked isoform composed of a 40 kD cytokine receptor-like subunit encoded by this gene and a 35 kD subunit encoded by IL12A. This cytokine is expressed by activated macrophages and is an important inducer of Th1 cell development. | |||||||||||||||||||||||||||||||||
| 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 | 3.12-200 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.2 ★★★★★
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Product Reviews
★★★★★ 5
Cute, lightweight and so roomy!
Size: Large, Color: Amourpink, Size: Large, Color: Amourpink
I am honestly so impressed with this backpack! It is the perfect mix of cute, comfy, and practical. The soft blush pink color is beautiful in person and gives it such a clean, stylish look without being flashy.
What surprised me most is how spacious it is. It holds SO much more than it looks like it would. I was able to fit all my travel essentials inside with room to spare, and the different compartments make it easy to stay organized. The side pockets are super convenient, and I love that it slides right onto my suitcase handle for traveling.
The straps are also really comfortable and padded, so even when the bag is full it still feels easy to carry. The material feels lightweight but durable, which makes it perfect for travel, work, day trips, or even everyday use.
If you want a backpack that is both functional and aesthetically pleasing, this one is definitely worth it. Cute, cozy-looking, lightweight, and surprisingly roomy!
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Reviewed in the United States on May 12, 2026
★★★★★ 5
Amazing product!! I use it so much
Size: Large, Color: Amourpink, Size: Large, Color: Amourpink
Absolutely LOVE this back pack I use it all the time! It holds a ton of stuff and has a lot of pockets and holds a good amount of stuff. It’s great for a simple sleep over and even flights. It’s very durable and great as a personal item on flights. The strap to keep it on your roller bag is great and it also has a ton of straps to adjust it
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Reviewed in the United States on May 24, 2026
★★★★★ 5
Spacious, organized, and perfect for travel!
Size: Large, Color: Black
This travel laptop backpack is absolutely fantastic! It's spacious and well-organized with multiple compartments for everything you need. It's airplane carry-on approved and fits perfectly in the overhead bin. The laptop compartment holds up to a 17" laptop securely with great padding. The water-resistant material keeps everything protected and the straps are comfortable for long trips. Great quality for the price. Highly recommend!
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Reviewed in the United States on May 26, 2026
★★★★★ 4
Good bag for a good price
Size: Large, Color: Amourpink
Really loved this backpack. I got it after reading all the reviews and wanting something for travel. The inside pockets with extra compartments that are water proof or organizing was a really nice touch. I traveled on airlines with this a few times and then started using it for school as well. It help up really well. I tend to be kind of rough on things and unfortunately the zipper broke. I'm unsure of if I over stuffed it or it was just a cheaper zipper. I would have considered resorting it because I used and loved it so much but my mom's dog got to it and it was not salvageable after that. I would recommend this specifically as a great travel backpack for the air port but it can easily double as an over night bag or school back pack.
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Reviewed in the United States on April 13, 2026
★★★★★ 5
So big, but lightweight!
Size: Large, Color: Amourpink
I bought this backpack to have as my "personal item" on the many flights I have to take. It is the perfect weight, having a place for just about anything you'd want to have during a flight, as well as having your carry-on items. The charging port is a bit of a challenge to plug in because of how you have to position the cord, but it is doable. I did not end up using the charging feature on my most recent flight, but I can see how it would be helpful for the travel that does not provide many charging stations. I have some serious back issues, but even full, I found this backpack to be a reasonable weight. I cannot attest to it's durability at this time, as I've only taken it on two trips now, but it is a great bag. There are many compartments, which can be good and bad. I recently put my wallet with all my identification and cards in a compartment that was easily missed. I thought I had lost my wallet and was panicked about how I'd get on my return flight if I didn't find it. After completely emptying the bag in my friend's car, and running my hands all over the compartments, I found the wallet. Here's the thing, my wallet is not little, so you can see that the walls of this backpack are pretty thick. The pink color is more of a dusty pink, which I find to be more attractive than a bright pink for something like this.
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Reviewed in the United States on May 22, 2026