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Description
Mouse TGFa 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. 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 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 Transforming Growth Factor Alpha (TGFa) 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 Transforming Growth Factor Alpha (TGFa) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Transforming Growth Factor Alpha ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Transforming growth factor alpha (TFG-α), a protein encoded by the TGFA gene, is a member of the epidermal growth factor (EGF) family and a mitogenic polypeptide. It can induce epithelial development and transform a variety of cells. Its amino acid sequence is similar to that of epidermal growth factor. It is produced by macrophages, brain cells, and epidermal cells. Similar to epidermal growth factor, it exerts its effects by binding to the epidermal growth factor receptor (EGFR), a receptor on the cell surface. Upon binding to this receptor, which mediates protein kinase activity for cellular signaling, the protein becomes activated. | |||||||||||||||||||||||||||||||||
| 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 | 15.62-1000 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.4 ★★★★★
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Product Reviews
★★★★★ 5
Very comfortable pillow
Size: Queen (1 Count), Color: White
I’ve been sleeping with this pillow for a few nights and I really like it. It’s soft but still supportive, and it stays cool during the night. I also like that you can adjust the filling to make it more comfortable. Definitely helped me sleep better.
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Reviewed in the United States on May 27, 2026
★★★★★ 4
Fantastic pillow for the price, adjustable is a different ball-game entirely.
Size: Queen (1 Count), Color: White
Overall, I'm quite pleased with this. The fact that it is fully adjustable (adding or removing foam, and they give you a bunch extra) is a massive boon as I like pillows to be pretty low profile. It feels nice to the touch, and compresses nicely but not a ton like down. The memory foam that is inside of it seems to be combined with some kind of a harder material, it's all shredded so I can't tell what it is, but it seems like maybe plastic or something of the kind. Hasn't been an issue in any way, but was just weird feeling it when removing foam.
I took a point off because there seems to be some false advertising going on. You can see in their picture that there is a tag that says bamboo side, the other side of that says cooling side. They also mention it in their advertising. As far as I can tell that is not true at all. The cover unzips so that you can remove the material so you can see that there is nothing on either side of it, and both sides appear visually identical to each other. Certainly I would NOT call this a cooling pillow in any way. It's maybe decently cool if you leave it in a cool place for a while, but over the course of the night warms up quite a bit. I did not notice any appreciable difference in the cooling of either side of it.
There is a definite noticeable smell to it, but I wouldn't call it 'bad' like most memory foam things are when they come from the factory. It smells kind of like some kind of cleaning solution in a noticeable but not unpleasant sense. All in all, for the price this is a fantastic pillow and I would definitely recommend it.
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Reviewed in the United States on January 14, 2026
★★★★★ 5
Most Comfy Pillow Ever Used
Size: King Size Pack of 2, Color: White
I recently purchased the BETU Cooling King Size Pillows Set of 2, and I have been thoroughly impressed with their quality and performance. These pillows have truly transformed my sleeping experience, making them an essential addition to my bedroom.
First and foremost, the shredded memory foam provides fantastic support while remaining incredibly comfortable. These pillows adapt perfectly to my head and neck, offering excellent support for back, neck pain relief, and side sleeping. The adjustability of the memory foam allows me to customize the firmness to my liking, ensuring a perfect balance between soft and firm support. This customization has made a tremendous difference in my sleep quality, reducing any discomfort and ensuring I wake up feeling refreshed and pain-free.
One of the standout features of these pillows is their cooling capability. As a hot sleeper, I struggled to find pillows that keep me cool throughout the night. The BETU Cooling Pillows effectively dissipate heat and maintain a comfortable temperature, allowing me to enjoy a restful and uninterrupted sleep. The cooling technology is truly a game-changer for anyone who tends to overheat during the night.
The pillow covers are another bonus. They are soft, breathable, and easily removable for washing, ensuring that the pillows remain fresh and hygienic. The white cover adds a clean and elegant look to the bedding, making them aesthetically pleasing as well.
The king size is generous and luxurious, providing ample space and comfort. Whether used for sleeping, lounging, or reading in bed, these pillows offer an unparalleled level of comfort and support.
In summary, the BETU Cooling King Size Pillows Set of 2 is an exceptional product. The combination of shredded memory foam, adjustable firmness, and cooling technology makes these pillows perfect for hot sleepers and those in need of superior support for back and neck pain. The high-quality covers and overall design further enhance their value. I highly recommend these pillows to anyone looking to improve their sleep quality and comfort.
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Reviewed in the United States on July 31, 2025
★★★★★ 5
Cooling pillow
Size: Queen (1 Count), Color: White
Good pillow very comfy and cool. Good quality material very durable. I love using it. Perfect size!
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Reviewed in the United States on June 5, 2026
★★★★★ 5
Cool, Supportive, and Great for Hot Sleepers
Size: Queen Size Pack of 2, Color: White
These cooling pillows have been a great upgrade for my sleep. The shredded memory foam provides nice support while still feeling soft and adjustable. I like that the pillows keep their shape and don’t go flat during the night.
The cooling fabric really helps if you tend to sleep warm. They feel breathable and comfortable, especially compared to traditional memory foam pillows that trap heat. The queen size is generous and works well for both side and back sleeping.
Overall, a great value for a set of two pillows. Comfortable, supportive, and noticeably cooler than my old ones. I would definitely buy them again.
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Reviewed in the United States on March 6, 2026