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Description
Rat IL-12 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. Preparation of 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 2000pg/mL). Then dilute to the following concentrations: 2000pg/mL, 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.25pg/mL, and 0pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each tube. Pipette 500uL of the 2000pg/mL standard working solution into the first EP tube and mix thoroughly to make a 1000pg/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube is used 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 Interleukin 12 (IL-12) 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 12 (IL-12) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Interleukin 12 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Interleukin-12 (IL-12) is 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 | 31.25-2000 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.8 ★★★★★
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Product Reviews
★★★★★ 5
Nutrient-Packed Spirulina Powerhouse!
I've been taking the Micro Ingredients Organic Spirulina Supplement, and it has exceeded my expectations in every way. This organic spirulina supplement is a true powerhouse of nutrients, and it has become an essential part of my daily routine.
One of the standout features of this supplement is its potency. With 3000MG per serving and 720 tablets in a single bottle, it offers exceptional value and a 4-month supply. That means I get all the benefits of spirulina without the need for frequent reordering.
I appreciate that these tablets are free from fillers and are Non-GMO. It's reassuring to know that I'm consuming a pure and clean source of spirulina.
The nutritional benefits are remarkable. Spirulina is rich in vegan protein, vitamins, and prebiotics, making it a fantastic addition to my diet. I've noticed an increase in energy and overall well-being since incorporating it into my daily regimen.
The convenience of tablets makes it easy to incorporate into my busy schedule. There's no need for mixing or blending, which is a time-saver.
In conclusion, the Micro Ingredients Organic Spirulina Supplement is a game-changer in the world of dietary supplements. It offers a premium source of organic spirulina, packed with essential nutrients and free from fillers. I highly recommend this supplement to anyone looking to boost their overall health and well-being. It's been a valuable addition to my daily routine, and I couldn't be happier with the results!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on September 20, 2023
★★★★★ 4
Real Happy with Purchase
Happy these are tablets instead of capsules.
Easier to take then dealing with the powder version of Spirulina.
No bad taste, or aftertaste.
They seem to do what they are supposed to for my health regimen.
Good price
I DO recommend this product
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Reviewed in the United States on February 9, 2026
★★★★★ 5
Great value for a great product
Great price for bulk spirulina. This is comparable tot he organic Sprouts brand I like. I only do 1 a day so this has lasted over a year!
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Reviewed in the United States on March 24, 2026
★★★★★ 5
Energy and value
Love these ! Great value for the money, big bag, and no taste or aftertaste at all! Easy to swallow and they give this 61 year old much needed energy to get more things done! I’m very impressed
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Reviewed in the United States on May 21, 2026
★★★★★ 5
Amazing Energy Boost
This gave me an energy boost immediately after I first took it. It worked so well, so fast, I thought it could not be from this product. Nothing works that fast right? But this did for me.
This is my first time taking Spirulina. I’ve taken supplements and used energy drinks but none had noticeable effects. This Sporulina does.
It is easy to take in this pill form. I curled my fingers in the photo to show how much green residue gets on you when you reach in the bag. It’s a bit more when the bag is brand new. I am halfway through the first, around two months. The green washes off easily with water and does not stain.
It does smell like fish. The odor is stronger when the bag is new. It barely has any taste so breathing through your mouth makes it easier to take if you don’t like fish. I like fish, but not strong fishy fish like mackerel. This is medium fishy when new and is still easy for me to take.
The energy boost was incredible for me. You might not have the same experience. I am a bit older than middle age male. That likely played into my increased sluggishness.
I sleep better after starting to take this. I no longer need to sleep in, and I am not foggy headed all morning. I’ve never been a morning person but now find I can hop out of bed and be ready to go early in the morning. Incredible for me.
I now fall asleep easier and stay asleep. When I do get up for the bathroom I can fall right back to sleep. Ive had sleep apnea my entire post HS life and possibly before. This did/will not cure that and I still use a CPAP. I was alway very trim and fit. So I wonder if this would have helped my sluggishness in my younger years. If that is you I would definitely give this a shot.
I have no idea what is in this that I needed but it is nothing short of amazing for me. Quality multi-vitamins or greens never did this for me.
Hard to beat all that and that a four month is just over $20. Hopefully the price stays there.
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Reviewed in the United States on July 17, 2021
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