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Description
Rat TGF-b2 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: 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: 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. 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. 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. 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. 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. 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 Beta 2 (TGF-b2) 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 amount of Transforming Growth Factor Beta 2 (TGF-b2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Transforming Growth Factor Beta 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Transforming growth factor β2 (TGF-β2), also known as glioblastoma-derived T-cell suppressor factor (G-TSF or BSC-1), is a secreted protein called a cytokine. It performs many cellular functions and plays a crucial role in embryonic development. It is an extracellular glycosylated protein and is known to inhibit the action of interleukin-dependent T-cell tumors. It regulates various processes, including angiogenesis and cardiac development. Its immune-regulating effects include: reducing inflammation: Through regulatory T cells, it helps balance the immune system and reduce inflammatory substances; reducing allergic reactions: It reduces the level of allergies caused by IgE (immunoglobulin E), reducing hypersensitivity; improving food tolerance: It helps build tolerance and mitigates the risk of allergies to foreign proteins in food; and protecting against viral and bacterial invasion: It promotes IgA (immunoglobulin A) secretion, which helps the intestinal mucosa develop and resists foreign invasion; and protecting against influenza A virus (H1N1) infection, reducing the severity and duration of the disease. | |||||||||||||||||||||||||||||||||
| 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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4.7 ★★★★★
Based on 260 reviews
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Product Reviews
★★★★★ 3
There are better books out there
Format: Paperback
I knew so much of what I read that I stopped reading 2/3 into the book. This is an average book.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 4, 2022
★★★★★ 5
Great book nice addition to howe library
Format: Paperback
The kid i bought it for loves it and rereads often. The paper is the nice shiny kind and colorful
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on September 13, 2025
★★★★★ 5
Another glimpse of the boy-behind-the-bat, with humor and heart; 4.5 stars
Format: Kindle
1st Line: "This is Gotham City."
Review: This new graphic novel, of a pre-Batman Bruce Wayne as a kid, opens in a Gotham City already all but decimated by crime and corruption. But with all its horrors, the worst of them for the young, rich, highly-intelligent but otherwise nerdy Bruce Wayne, orphaned after the murders of his parents, is none other than the Gotham Preparatory School for the Really, Really Gifted - his middle school. Here, all the students have some form of super power or another (The Flash, Wonder Woman, Superman, Penguin, Catwoman, and more - all started off in middle grade here) ... except for Bruce, who only got into the school because his parents funded the building of it before their deaths, and remains a target for bullies for not having any powers of his own. But when fellow student Jack Napier, himself having no super powers except for conning his way into the school, begins bullying Bruce's friend, elementary student Dick Grayson, Bruce becomes determined to strike back - thus beginning his ascent to a career as a vigilante, even as his loyal butler Alfred and the school's vice-principal forbid it. But will Bruce get it together in time to foil Jack's plans for the Crime of the Semester? And what of Bane, Jack's new sidekick, who would be a formidable foe even for someone with superpowers?
Bruce Wayne: Not Super is terrific; a comedic look at the young, super-smart yet geeky Bruce completely out of his element as a young kid, formerly with the attention span of a goldfish, who may have finally found his calling. Also nice are the glimpses of a number of future DC superhero icons in their youth, but the story really does belong to the humor of Bruce wanting to save his city, agreeing to take a hyperactive Robin in tow as his sidekick, with no knowledge of how to fight and an arsenal of weapons thrown together in minutes - and potentially of harm more to the user than the bad guy - all while Alfred says no and remains in constant threat of sending the future Batman to his room. The poor kid, after all, can't even put a bat costume together; in it, he's mistaken for everything from a badger to a rat to even a prairie dog. Bruce pushes past it all, his pluck and determination to save Gotham City one villain at a time his true calling, and among writer Stuart Gibbs glib and funny humor and Berat Pekmezci's bursting-with-color art, the very basic but heartfelt beginnings of the boy who'd grow to become The Bat shines through. It's really good. So good, I already itch for a sequel. 4.5/5 stars
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Reviewed in the United States on March 14, 2023
★★★★★ 4
Fun story with familiar characters
Format: Paperback
4 stars = Great! Might re-read.
This was fun! I enjoyed the illustrations and getting to scan the crowd scenes for more DC characters. I enjoyed Bruce's journey. I didn't love the portrayal of Clark Kent here, but the rest of this was fun. I would definitely read more of these if there are going to be sequels.
There's a one-star review here that is completely on target in their assessment of some of the liberties taken in the book. Buyer/reader will have to decide if those observations will lead you to skip this one or if you can set aside those internal inconsistencies to accept the story as is.
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Reviewed in the United States on April 15, 2023
★★★★★ 5
Fun for kids and those kids at heart!
Format: Paperback
When I was a kid around 8 or 9 years old, we would go to an outdoor theatre, a special treat. I remember the old 1960's era Batman movies playing on the screen. It was fun, exciting, heroic...all those things that made it pure enjoyment. When I think of recent Batman movies that have turned dark, deadly, desperate, all the fun has gone out of those experiences and I simply don't watch those movies. This is simply a long way of saying that this book, Bruce Wayne: Not Super, brought back all the fun I experienced as a kid. Here we have our hero, who doesn't think he is, who compares himself to everyone around him with supernatural abilities, and begins to grow desperate. The background story is essentially the same: dead parents, rotten town, etc. But we see life from his perspective and can root for him all the way. He goofs up, makes lots of mistakes and this makes him even more lovable. This book is prefect for readers young and old, and if this made into a movie, I guarantee you that I would watch it. Highly recommended.
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Reviewed in the United States on April 4, 2024
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