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Description
Human E2F1 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 enzyme conjugate are sequentially added to microwells pre-coated with a transcription factor E2F1 (E2F1) capture antibody. After incubation and washing, the sample 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 transcription factor E2F1 (E2F1) content in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Transcription factor E2F1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | The transcription factor E2F1, also known as E2F1, is a protein encoded by the E2F1 gene. The protein encoded by this gene is a member of the E2F transcription factor family. The E2F family plays an important role in controlling the cell cycle and tumor suppressor gene function and is also a target of small DNA oncogenic viral transforming proteins. Multiple members of the E2F family contain many evolutionarily conserved domains. This protein, like the other two members, E2F2 and E2F3, possesses an additional cyclin-binding domain. | |||||||||||||||||||||||||||||||||
| 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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4.6 ★★★★★
Based on 2368 reviews
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Product Reviews
★★★★★ 5
Peanut butter delivery system
Color: Blue
My big dog loves this bone. I run PB down the middle on one side when we go out. He loves it and brings the bone to us when we return to play :gimme that bone.”
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Reviewed in the United States on June 3, 2026
★★★★★ 4
Durable bone that you can add a spreadable treat to
Color: Blue
Our German Shepherd was 4 months old when we purchased this. She’s going to be 6 months soon. She has been teething really bad. We wanted some way to sooth her pain. This product was just what we needed. We put it in the freezer with some peanut butter in the center on both sides. Once frozen we give to our puppy. She loves it. When we open the freezer she recognizes the ziplock package that it was delivered in which we store it in our freezer. She jumps around until we give it to her. She loves it and it keeps her occupied for a while. It is very durable. I did not notice any smell when we received it. It does not make any noise. The plastic material is safe for animals. To clean I put in the dishwasher. The only reason I did not give it 5 stars is because sometimes the peanut butter gets stuck in it and may need to go through the dishwasher twice which is not a big deal. Or you can hand wash it. I found the dishwasher was best for us. You do not even have to put anything on it, but our pup likes it. I would recommend to others.
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Reviewed in the United States on June 15, 2025
★★★★★ 5
My dog likes them
Color: brown
Dogs play with these and chew on them
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Reviewed in the United States on June 12, 2026
★★★★★ 5
Highly recommended for any owner of a "Super Chewer."
Rating: ⭐⭐⭐⭐⭐ (5/5)
Title: Finally, toys that can actually withstand my Labrador’s jaws!
If you own a "power chewer," you know the struggle: you bring home a new toy, and within ten minutes, it’s shredded into a thousand pieces of foam and plastic. After going through countless toys that promised durability but failed within a day, I decided to give the WOWBALA 2-Pack Indestructible Dog Toys a try—and I am genuinely impressed.
The Durability Factor
The biggest selling point here is the material. These are made from high-density, food-grade nylon, and you can feel the quality the moment you take them out of the package. They are solid, heavy-duty, and have held up perfectly against my Lab’s relentless chewing. Even after a week of heavy use, there are only minor surface marks where his teeth have made contact, rather than the dangerous chunks that usually break off other toys.
Design & Engagement
The pack comes with two different shapes—a stick and a ring—which is a great variety. The curved, ergonomic design of the stick makes it very easy for my dog to hold between his paws while he chews, and the textured surface seems to really hit the spot for him. I’ve also noticed that it helps keep his breath fresh and his gums healthy since the texture acts almost like a toothbrush while he plays.
The "Milky" Scent
One thing I was initially skeptical about was the "milky scent." I wondered if it would be overpowering, but it’s actually quite subtle. It was strong enough to immediately pique my dog’s curiosity right out of the box, but it doesn't leave a weird smell on my carpet or hands. It’s clearly effective, as he keeps coming back to these toys even when he has others scattered around the room.
Peace of Mind
Beyond just being tough, I appreciate that they are non-toxic and dishwasher safe. After a long day of outdoor play in the mud, being able to just toss these into the dishwasher and have them come out clean and sanitized is a huge convenience.
Final Verdict
If you have a medium to large breed dog that loves to destroy everything in sight, these toys are well worth the $15.99 investment. It’s rare to find a product that lives up to the "indestructible" label, but these truly do. They’ve been a lifesaver for keeping my dog occupied while I’m working from home, and it’s a relief not having to play "clean up the debris" every evening.
Pros:
Extremely durable high-density nylon.
Great value (2-pack).
Easy to clean (dishwasher safe).
Engaging design for teething, training, and boredom relief.
Safe and non-toxic.
Cons:
Because they are hard nylon, be careful using them indoors on hardwood floors—they can be a bit noisy when dropped!
Highly recommended for any owner of a "Super Chewer."
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Reviewed in the United States on May 19, 2026
★★★★★ 5
These are way better than I expected and hold up to our aggressive chewer spectacularly!
I got these specifically for our newest pup (an 8 month old lab/pitbull mix--she's a big puppy, lol) who is obsessed with toys and chewing. She has a tendency to go through her toys quick, both chew toys and soft ones, so I wanted something that would last a bit longer for her because I can tell she gets a bit sad when we have to trash one that's hit its lifespan. So far, these have been a big hit! They're the perfect size for her to pick up, the chicken claw-style texture is great for both keeping her interested and giving her some grabbing options, and they do seem to hold up to their claim of being made for aggressive chewers! There's obviously some wear, but it looks like expected chew toy wear; no claws have snapped off, and her chew marks look relatively minimal compared to what she's done to other chews. I also appreciate that these came in a two pack; we have a smaller dog, and while these are technically big for him, he enjoys having one to play with and chew on as well. These have been a pleasant surprise and I actually think I'll come back to buy them again whenever they're spent! Definitely a worthy purchase!
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Reviewed in the United States on June 2, 2026
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