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Description
Mouse TGFb3 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 culture supernatant: Centrifuge at 1000×g for 20 minutes, take the supernatant for detection, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. 5. 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 Transforming Growth Factor Beta 3 (TGFb3) 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 Beta 3 (TGFb3) 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 Beta 3 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Transforming growth factor beta-3 is a protein encoded by the TGFB3 gene. It is a type of protein, known as a cytokine, involved in cell differentiation, embryogenesis, and development. It belongs to a large family of cytokines known as the transforming growth factor beta superfamily, which includes the TGF-β family, bone morphogenetic proteins (BMPs), growth and differentiation factors (GDFs), inhibins, and activins. TGF-β3 is thought to regulate molecules involved in cell adhesion and extracellular matrix (ECM) formation during palate development. Without TGF-β3, mammals develop a malformation known as cleft palate. This is caused by the failure of epithelial cells on both sides of the developing palate to fuse. | |||||||||||||||||||||||||||||||||
| 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 | 0.15-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell culture supernatant and other biological fluids |
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4.8 ★★★★★
Based on 30 reviews
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Product Reviews
★★★★★ 5
10/10 Great game changer for aggressive chewers
After a scary experience where our Doberman ingested part of a toy and needed emergency surgery, we had to get rid of all plush toys and small balls in our house. Replacing everything felt overwhelming.
This ball has been fantastic. It’s truly built for aggressive chewers and holds up incredibly well. It’s perfect for fetch, strong enough for tug-of-war, and the bounce is excellent, which keeps our dog fully engaged during self-play when he’s entertaining himself. That unpredictable bounce makes it exciting without being unsafe.
Most importantly, it gives us peace of mind. No shredding, no chunks coming off, no felt cover and constant worry about ingestion. Since having to replace all of his previous toys and balls, this one has stood out as the best by far.
If you have a large, powerful dog—especially a Doberman—or a dog who destroys toys quickly, this is a must-have. Durable, safe, and genuinely fun. 10/10, highly recommend.
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Reviewed in the United States on January 21, 2026
★★★★★ 5
Finally, a sturdy oversized ball my boy can't destroy! And can pick up easily
This is just wonderful! My dog has destroyed all other balls! I think this is finally one he cannot destroy. And I love the holes so he can pick it up and throw it himself. LOL. And it BOUNCES, bringing him endless joy and skill in catching it. Cause you don't know which way it will bounce off the ground. Oversize was just right for him. Glad to have found this great sturdy ball.
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Reviewed in the United States on April 29, 2026
★★★★★ 4
Almost perfect...upraised logo should be left off
This toy is great for relentless chewers with strong jaws! It would be almost perfect if it didn't have a "K9" logo in raised rubber on it. That is the place that my aggressive chewer shepherd/bulldog mix focuses on and is able to get small chunks off. Without that logo, he would chew the ball more evenly, instead of fixating on that one spot. He loves this ball and has chewed on it for about an hour off & on each of the 11 days he has had it. Although it is very durable so far, the flaw is the upraised logo.
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Reviewed in the United States on January 22, 2026
★★★★★ 5
Perfect for High-Drive Dogs & Surprisingly Durable (six month review)
I use this ring ball for structured fetch and tug sessions with my high-drive German Shepherd mix (91 lbs. at 10 mon.), and it’s easily the best toy we’ve added to our routine. What makes it truly exceptional is how it allows my dog to engage in the full canine predatory sequence:
• Chase: He locks in visually and bolts after the ring when I throw it.
• Grab & Hold: He brings it back, pins it against my leg, and bites down firmly (the toy—not the leg!)—his way of acting out the natural catch-and-control behavior.
• Unalive & Tug: We finish with a strong tug session, which satisfies his instinct to grip and shake—basically his version of “making the toy unalive.”
• Settle & Chew: Afterward, he relaxes with a chew toy or eats—completing the full physical and mental cycle in a healthy, balanced way.
Durability is outstanding—this toy stands up to intense tug sessions with no tearing or damage, even with a 91 lb powerhouse. My senior dog also joins in for double tug play, and it holds up beautifully to that too.
I don’t leave this toy down for chewing—it’s not a chew toy. I use it specifically for fetch and tug, then put it away when we’re done. This keeps it exciting and extends its lifespan. My dog has other chew toys he can use freely throughout the day.
This isn’t just another fetch toy—when thoughtful it provides a structured, instinct-satisfying experience that’s mentally enriching and physically engaging. If you have a high-drive or working breed, or a dog who thrives on interactive play, this ring ball is 100% worth it.
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Reviewed in the United States on March 30, 2025
★★★★★ 5
Love these!!!
Color: Black and White, Color: Black and White
I absolutely love these socks!!! I use them so I can walk around the house when I put lotion on my feet. They are perfect. Very very smart young lady that came up with this idea.
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Reviewed in the United States on February 14, 2026
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