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Description
Mouse IL12 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. 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. 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. 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. 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. 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 1mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 1000pg/mL). Then dilute to the following concentrations: 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.25pg/mL, 15.625pg/mL, and 0pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 1000pg/mL standard working solution into the first EP tube and mix thoroughly to make a 500pg/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 12 (IL12) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of Interleukin 12 (IL12) 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 12 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Interleukin-12 (IL-12) is an interleukin naturally produced by dendritic cells, macrophages, neutrophils, and B lymphocytes (NC-37) in response to antigen stimulation. IL-12 belongs to the interleukin-12 family and plays a key role in the activity of natural killer cells and T lymphocytes. IL-12 also exhibits anti-angiogenic activity, meaning it can inhibit the formation of new blood vessels. IL-12 is a disulfide-linked heterodimer composed of two subunits: a 40 kDa (p40) and a 35 kDa (p35) glycoprotein. | |||||||||||||||||||||||||||||||||
| 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 | 15.6-1000 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.2 ★★★★★
Based on 26 reviews
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Product Reviews
★★★★★ 5
Fun for an active dog!
I have an Australian Shepherd that absolutely loves this auto rolling dog ball! The ball is hard, but covered in a rubber coating. It is still loud when it hits the floor, but that is to be expected. My dog has chased this as well as thrown this ball, and it is still working flawlessly! This ball is definitely durable! It is easy to charge, and provides hours of entertainment! The ball is a great size for a medium dog. I recommend it!
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Reviewed in the United States on June 5, 2026
★★★★★ 1
Auto-Rolling Dog Ball - loud with a mystery filling.
I got this to entertain my high energy dog and she does like it, but I don't.
The ball is incredibly loud and doesn't move around nearly as much as I had hoped. When your dog picks it up, it turn their mouth into an amplifier making it louder.
It bounces/vibrates around more than rolling. Which is not what I expected, but seems to be engaging enough for more dog (but that doesnt take much).
What I really dislike is there is no way on the exterior to know what mode you are in and the inside is full of grease. To charge it and to see the lights to identify what mode it is involves opening it and touching the grease covered insides. The lights don't seem to match the instructions when trying to switch modes. The light is supposed to stay on for 3s. It doesn't for either mode and immediately cycles through other colors making it more confusing.
The exterior seems durable enough for a moderate chewer and I like the color and texture.
At $15, it seems like a fair price, but I can't recommend this at any price point. I don't feel comfortable letting my dogs play with toys that could easily expose them to an unidentified grease. Any greased components should be sealed. Not somewhere I have to touch for normal operation and potentially leak.
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Reviewed in the United States on April 4, 2026
★★★★★ 5
Gentle vibrations and rolling
My sweet old man dog didnt know what to think of this at first. We introduced it to him without turning it on at first, like a regular ball. That was fine, he was good with that and he liked to go chasing it (its heavy because its mechanical so I wouldnt throw it, a light toss/roll is how we started).
After we played that way a while we turned it on. At first he was confused as to why his toy was chasing him. It mostly vibrates and bops around a bit. But then my pup started chasing it and kicking it around like a little soccer ball and he LOVED THAT. A charge lasts quite a while too and if he doesnt play with it in for a few minutes it goes to sleep so its not just bouncing constantly and driving me crazy.
My pup likes it and it gives me a rest from the constant throwing so I'm happy!
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Reviewed in the United States on April 7, 2026
★★★★★ 5
Keeps My Dog Entertained for Hours!
This auto-rolling dog ball has been a total game changer for keeping my dog busy and active! The second I turned it on, my dog was instantly curious and started chasing it all over the house. It moves and rotates in a really fun, unpredictable way that keeps their attention much longer than regular toys.
I love that it helps with boredom relief, especially on days when we can’t get outside as much. It’s perfect for indoor play but also works great outside too. The material feels durable and holds up well against chewing, which is a must for my pup.
The USB rechargeable feature is super convenient—no constantly replacing batteries—and it holds a charge long enough for multiple play sessions. I also appreciate that it’s easy to clean and has some water resistance, so I don’t have to worry too much about slobber or a little outdoor use.
Overall, this is a fun, interactive toy that actually keeps dogs engaged and moving. Highly recommend for anyone with an energetic or easily bored pup!
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Reviewed in the United States on April 5, 2026
★★★★★ 5
Great toy!
Pattern Name: Carrot
"Dog toys by Dole? The banana people??" Yep, me too. My dog loves this toy. The plastic piece is a great texture for chewing. The stuffy is small enough for most puppies and will grow to medium sized dogs. The green of the carrot crinkles and they both wash up good. The stuffy has been through the wash many times. Good quality, durable toys.
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Reviewed in the United States on June 4, 2026
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