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Description
Mouse IRG1 ELISA KitProduct Specification Usage Required experimental equipment: 1. Microplate reader (450nm) 2. High precision pipettes and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and
Product Specification
| Usage | Required experimental equipment: 1. Microplate reader (450nm) 2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37°C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL 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 collect the supernatant for analysis. Cell Lysis Buffer: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended 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 freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. 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 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 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 an immunoreactive gene 1 (IRG1) 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 immunoreactive gene 1 (IRG1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Immunoresponsive gene 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Immune response gene protein 1 (IRG1) is a metabolic enzyme in the tricarboxylic acid cycle that catalyzes the decarboxylation of aconitic acid to itaconate. It encodes a protein involved in pathways including glucose and energy metabolism. IRG1/itaconate is a crucial link between macrophage immunity and metabolism. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, and other biological fluids |
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4.0 ★★★★★
Based on 1083 reviews
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Product Reviews
★★★★★ 4
Built-in USB-C solves the power issue, but it is strictly for moderate chewers
Color: Blue
Got this to keep the high-energy puppy occupied indoors so the older dogs can actually get some peace and quiet. The main issue with most automated electronic pet toys is that they rely on expensive, hard-to-find button cell batteries that die after a single afternoon of use.
This unit utilizes a built-in 300mAh battery with a standard Type-C charging interface. That is a mandatory design feature if you actually want to run the toy daily without going broke replacing power cells. The internal motor generates enough erratic movement to keep a dog's attention, and it actually has the torque to navigate over standard tile, hardwood, and light carpets without instantly stalling out in a corner.
You need to completely understand the physical limitations of what you are unboxing here. The listing explicitly states this is not for heavy chewers, and you need to take that warning seriously. The outer shell is TPU, but the internal chassis housing the electronics is hard ABS plastic. If you hand this to an 80-pound working dog that aggressively shreds thick bones, they are going to crush the housing and destroy the internal motor assembly in about ten minutes.
At roughly 2.5 inches in diameter, it's basically the size of a standard tennis ball. If you use it as intended—as an interactive chasing toy for a puppy or a moderate chewer rather than a heavy-duty chew toy—it functions perfectly. It is a highly practical piece of indoor enrichment gear. Solid return on investment for burning off excess canine energy.
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Reviewed in the United States on May 29, 2026
★★★★★ 5
Pawsitively Stimulating🐾
Color: Blue
When my hooman brought this home, I thought it was fur their computer. After all, it has a plug and lights up. When they placed it on the floor, I was pawsomely surprised. Fur real, this little ball is wierd. It doesn't taste at all like my favorite tennis ball. I was a pawplexed that sometimes when I was finally brave enough to get it and hold it in my mouth, it would start jumping around. I think my hoomans finally found a toy that will keep me busy for a while. Mom is howling with excitement because it kept me interested for a long time. I hope I can sleep tonight. I see mom has plugged in my new toy to keep it "charged"- whatever that means....I never had a toy that came with its own leash before. Mom says she is gonna save the toy until other hoomans come over to keep me from being too ruff with them. You see the video, do I look a dog who could be too ruff?- Sincerely, the family dog, a bona-fido toy reviewer
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Reviewed in the United States on May 3, 2026
★★★★★ 4
Freaks my dogs out.
Color: Blue, Color: Blue
My dogs really don't know what to make of these. They mostly follow it around, staring at it curiously, but semi-afraid to approach it. That, or they'll bark at it. It charges quickly and has multiple modes to engage your pet with. It also locks closed securely ... for the most part.
That said, there are some downsides. This toy is mostly made for smaller dogs. I'd say 40 or 50 pounds max. While the plastic shell feels hard and durable, it's not hard enough for a big dog's jaws. My big bulldog mix bore down on this with his teeth, and I hard the plastic starting to snap. It's tough, but not great if your dog is the type who likes to put the pressure on.
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Reviewed in the United States on May 4, 2026
★★★★★ 5
Best money for a dog toy I've spent in a long time
Color: Blue
This interactive ball is small but packs a lot of fun and playtime for my dog's they go crazy every time we bring it out and turn it on, it lights up has several modes vibrates they love it will be purchasing amount for certain
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Reviewed in the United States on May 9, 2026
★★★★★ 4
Fun, engaging dog toy that keeps pets entertained, but battery life could be better
Color: Blue
I’ve been using the ITEHZTO Interactive Dog Ball Automatic Dog Toy for my dog, and overall it’s been a great way to keep him active and engaged, especially when I’m busy or not able to play fetch. The different play modes are a nice feature, and the unpredictable movement really grabs his attention. It does a good job of encouraging exercise and mental stimulation.
The USB-C charging is convenient, and I appreciate not having to mess with batteries. The size is also good for small to medium dogs, and it feels durable enough to handle regular play.
The reason I’m giving 4 stars instead of 5 is that the battery life doesn’t last as long as I expected, especially when using the more active modes. Also, on harder flooring it can be a bit noisy as it bounces around.
Overall, it’s a fun and useful interactive toy that my dog genuinely enjoys. With slightly longer battery life and quieter operation, it would easily be a 5-star product.
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Reviewed in the United States on May 6, 2026