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Description
Mouse EGR1 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 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 an Early Growth Response Protein 1 (EGR1) capture antibody. After incubation and washing, the assay is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of Early Growth Response Protein 1 (EGR1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Early Growth Response Protein 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Early growth response protein 1 (EGR1), also known as zinc finger protein 268 (ZIF268) or nerve growth factor-induced protein A (NGFI-A), is encoded by the EGR1 gene. The protein encoded by this gene belongs to the EGR family of Cys2His2-type zinc finger proteins. It is a nuclear protein that functions as a transcriptional regulator. The products of the target genes it activates are required for differentiation and mitosis. It is a tumor suppressor gene. It has a unique expression pattern in the brain, and its induction has been shown to be associated with neuronal activity. It plays a role in neuronal plasticity and is an important transcription factor in memory formation. It plays a crucial role in epigenetic reprogramming of brain neurons. | |||||||||||||||||||||||||||||||||
| 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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4.9 ★★★★★
Based on 30 reviews
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Product Reviews
★★★★★ 5
Very impressive quality
Color: Blue Octopus
An absolute HIT
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 11, 2026
★★★★★ 5
Perfect for "Heavy Chewers," lasts long, and has yet to come apart or tear up
Size: Large, Product Packaging: Standard Packaging
I have several dogs, and they are what would be considered as "Heavy Chewers." They are truly awesome pets, and I consider them like my children.
I started looking for toys that I wouldn't worry about if I didn't have line of sight for every minute. All dog toys, regardless of their size, type, weight, or if they're steel versus plastic material have a warning on the bottom of very label/package that states that the animal should never be left alone with the toy.
With that said, I do not leave the toys unattended, even though this toy has paid it's weight in how well it has stood up to the constant chewing my dogs give it everytime I place it in their crates to prevent fights, or issues on the pack order.
As far as the actual toy, it looks, feels, and even smells like a tire, but without any problems with the dogs accepting it to play with.
The toy bounces like a ball, and the overall weight is heavy enough to ensure durability. Just like a tire, the toy is made of rubber, not plastic material.
I have found that smearing small amounts of peanut butter in to the crevices of the tire rather than brand specific treats is just as effective, I will also freeze the peanut butter into the tire, and this also increases the length of time it takes for the dogs to consume.
I ended up buying three more toys, mostly so all my dogs have one, and that I also have one as a spare.
I would recommend this to any owner of a "Heavy Chewer" as the product is well made, and yet to have any defect or piece removed by any of my dogs, and they do chew on these toys extensively.
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Reviewed in the United States on April 13, 2025
★★★★★ 5
Fantastic dog toy
Size: Large, Product Packaging: Standard Packaging
Fantastic dog toy, probably my dogs favorite. It’s really durable, my dogs a heavy chewer and this toy has lasted several weeks. Will definitely buy again
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Reviewed in the United States on May 25, 2026
★★★★★ 5
Tough and Long-Lasting Toy for Heavy Chewers
Size: Large, Product Packaging: Standard Packaging
The KONG Extreme Tires are built for dogs who chew through just about everything else. Made from KONG’s most durable rubber, the toy is designed to stand up to strong jaws while also offering a fun shape that dogs enjoy rolling, chasing, and chewing. The tire design has grooves that can be stuffed with treats or peanut butter, which helps keep dogs engaged for longer play sessions.
This toy is especially useful for large breeds or heavy chewers who usually destroy softer toys in minutes. While no toy is completely indestructible, the Extreme Tires hold up far better than most and provide both chewing satisfaction and mental stimulation. The size is hefty and best suited for medium to large dogs, and smaller breeds may find it too heavy.
Pros: The rubber is extra durable, the tire shape adds variety and fun, and the grooves can be filled with treats for extra engagement.
Cons: It may be too heavy for smaller dogs, strong chewers can still wear it down over time, and it isn’t designed to bounce or fly like other KONG toys.
A rugged and reliable toy made for power chewers. The Extreme Tires provide long-lasting fun and double as a great option for treat-stuffing, making it one of KONG’s toughest and most useful toys.
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Reviewed in the United States on August 30, 2025
★★★★★ 5
Strong toy
Size: Large, Product Packaging: Standard Packaging
Nice, but smaller than I expected.
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Reviewed in the United States on May 16, 2026