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Description
Human TRA2b 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, biotinylated detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with a capture antibody against Transformer-2 protein homolog beta (TRA2b). After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of Transformer-2 protein homolog beta (TRA2b) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Transformer-2 protein homolog beta ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Transforming factor 2 beta (TRA2b), also known as splicing factor arginine/serine-rich 10 (SFRS10), is a protein encoded by the TRA2b gene. This gene encodes a nuclear protein that functions as a sequence-specific serine/arginine splicing factor, playing a role in mRNA processing, splicing patterns, and gene expression. It activates splicing of MAPT/Tau exon 10. It alters pre-mRNA splicing patterns by antagonizing the actions of splicing regulators such as RBMX. It binds to the AG-rich SE2 domain of SMN exon 7 RNA. It binds to pre-mRNA. Diseases associated with it include frontotemporal dementia and spinal muscular atrophy. It has been shown to interact with RBMX. | |||||||||||||||||||||||||||||||||
| 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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4.5 ★★★★★
Based on 25 reviews
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Product Reviews
★★★★★ 5
A perfect start
Format: Kindle
This is exactly the kind of graphic novel I was looking for. Full of sweetness with a little bit of bite. Reminds me of the great comic series bone. Great story, great art, I can’t wait to get into the next chapter.
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Reviewed in the United States on May 8, 2026
★★★★★ 5
Draws You in to the Series
Format: Paperback
The Stonekeeper is the first volume in Kazu Kibuishi's graphic novel series. Although aimed at middle school, it is good for upper elementary school children, and is enjoyable for adults as well. 2 years after the death of Emily and Navin's father in a tragic car crash, Karen moves her children to the family's old house in a dense forest. Built by Karen's grandfather Silas who then disappeared, it has been abandoned for years. During the first night, they hear a strange noise, and Karen is captured by an Arachnopod, a creature from a parallel world, Alledia. Emily and Navin follow their mother into the parallel world where they find their ailing great-grandfather Silas. He tells Emily about the power of the amulet she found in his house. It is the key to ruling Alledia. Silas expires, but not before instructing his robot and animal minions to aid Emily and Navin. This first volume, besides setting the story arc, narrates the rescue of Karen. Unfortunately, she is poisoned setting up the next volume for finding the antidote.
Besides an engaging story with compelling characters, Kibuishi is a fantastic illustrator. As with many exemplar graphic novels the spare text is amplified and much of the story told through his illustrations. The creatures of Alledia are strange enough to pique our imagination, but not so much that they are totally alien in form.
After reading Stonekeeper, I am eager to see what happens next with Emily and Navin, and their company.
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Reviewed in the United States on March 1, 2014
★★★★★ 5
wow
Format: Paperback
I remember when I read in reviews that kids had gotten sucked into this story and had become fans of this series so quickly.... Well, my daughter resisted till page 13 or 14... but since then she's in it... She's very picky when it is about books and even that she's 9, she started to read in English only a few months ago ( we lived abroad,she reads other language) so she still prefers books that is easy to read.
Anyway, get ready, this series is addicted... gooooood :) that is what we need :)
I mean, you can complain that this is a comic book but still, at least the child has the book in his/her hand and not a tablet... and at least he/she reads...
I'm really happy that I got this book for her.
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Reviewed in the United States on November 16, 2023
★★★★★ 5
Wonderful Story, But Small Print
Format: Paperback, Format: Paperback
My son really likes this book. He usually doesn’t enjoy reading much, but this one caught his interest right away. The illustrations are great, and the story is engaging too. He finished the book in just two days! I already bought the second one and am thinking about getting the whole series.
The only thing I didn’t like was the size of the letters. It wasn’t very easy for my 9-year-old to read on his own, and sometimes I had to help him.
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Reviewed in the United States on October 14, 2025
★★★★★ 5
Great story and great quality!
Format: Hardcover
These books are always great! One of my son’s faves. Great story and great quality as usual!
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Reviewed in the United States on May 15, 2026