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Description
Rat TAK1 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 conjugate are sequentially added to microwells pre-coated with a capture antibody against transforming growth factor-beta-activated kinase 1 (TAK1). After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of transforming growth factor-beta-activated kinase 1 (TAK1) 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-activated kinase 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Transforming growth factor kinase (TAK1), also known as mitogen-activated protein kinase kinase 7 (MAP3K7), is an enzyme encoded by the MAP3K7 gene. The protein encoded by this gene is a member of the serine/threonine protein kinase family. This kinase mediates signal transduction induced by TGF-beta and morphogenetic proteins (BMPs) and controls various cellular functions, including transcriptional regulation and apoptosis. It is a central regulator of cell death and is activated by a range of diverse intracellular and extracellular stimuli. It regulates cell survival not only through NF-κB but also through NF-κB-independent pathways, such as oxidative stress and receptor-interacting protein kinase 1 (RIPK1) kinase activity-dependent pathways. In response to IL-1, this protein forms a kinase complex that includes TRAF6, MAP3K7P1/TAB1, and MAP3K7P2/TAB2. This complex is required for the activation of nuclear factor kappa B. This kinase also activates MAPK8/JNK and MAP2K4/MKK4, thus playing a role in the cellular response to environmental stress. | |||||||||||||||||||||||||||||||||
| 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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4.9 ★★★★★
Based on 13 reviews
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Product Reviews
★★★★★ 5
Nice Suit For The Price
This suit was a time-saver! My wife ordered a tux that wasn't going to make it for a formal occasion, so this was ordered and came within three days. It was a good thing she purchased 2, in case of sizing errors. The extra large pants worked, but sizing down to the large jacket & vest made the best combination/fit. We didn't go with the blue/black bow tie & flower it came with, but may for another event. However, being able to work interchangeably at a low price was rewarding!!
Thought to add comments that the look of this suit is pleasing (though it's not the best thread). The workmanship gives it a 5 and even greater. The compliments were many. I do suggest using a good steam iron that will eliminate the wrinkles, without creating unwanted creases, while barely touching the suit; but, definitely follow the creases in the pants leg.
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Reviewed in the United States on March 20, 2025
★★★★★ 5
Disappointed
Size: XX-Large, Color: Light Grey, Size: XX-Large, Color: Light Grey
Overview of the Purchase Experience
I recently ordered a 3-piece suit for my significant other, who is quite tall at 6’8”. I was excited about this purchase, hoping it would be a stylish addition to his wardrobe. However, upon receiving the suit, I encountered several issues that I feel compelled to share.
Color Discrepancy
One of the most disappointing aspects of this order was the color inconsistency. The suit was supposed to be a cohesive set, but it arrived with two different colors. This mismatch not only detracted from the overall aesthetic but also made it difficult to wear as a complete outfit. When purchasing a suit, especially a three-piece one that includes trousers, a vest, and a jacket, one expects all pieces to match in color and style. Unfortunately, this was not the case here.
Quality Concerns
In addition to the color issues, I found the quality of the fabric and construction to be subpar. The material felt cheap and did not have the durability or elegance one would expect from a suit. The stitching appeared uneven in places, and there were loose threads that raised concerns about its longevity. A well-made suit should exude quality and craftsmanship; however, this product fell short on both counts.
Fit Issues Related to Height
Given my significant other’s height (6’8”), finding suits that fit well can be challenging. While I cannot recall his exact weight at this moment, it’s important to note that tall individuals often require tailored fits or specific sizing options that accommodate their stature. Unfortunately, even if the colors had matched and the quality had been better, I doubt this suit would have fit him properly without additional alterations.
Conclusion
Overall, my experience with this 3-piece suit has been disappointing due to color discrepancies and poor quality construction. For anyone considering purchasing this item for someone tall or for themselves, I would recommend looking elsewhere unless you are prepared for potential issues with fit and appearance.
Peggy Cole FabFashiion not approved
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Reviewed in the United States on September 24, 2024
★★★★★ 5
Excellent Affordable Suit for Special Occasions
Size: Large, Color: Black, Size: Large, Color: Black
Bought this for prom and was very happy with how it looked. The jacket is a nice medium weight and looked great on. We ordered a large, which was slightly big, but after having the suit tailored it fit perfectly. My son received many compliments and looked amazing.
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Reviewed in the United States on May 16, 2026
★★★★★ 4
Worked out perfect. Great value, quality suit
Size: Small, Color: Black, Size: Small, Color: Black
Worked out perfectly for my son’s 8th grade formal. Pictured he is wearing a men’s small. Quality but a little baggy. It fit fine but we had the jacket altered to fit tighter around his back and the pants were hemmed and tapered. Great value for the money. Does not include a white shirt. Includes jacket, vest, pants, bow tie and pocket pin. 4 stars only because of baggy but some may like that fit.
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Reviewed in the United States on May 4, 2025
★★★★★ 5
Awesome Suit
Size: Medium, Color: Black
Awesome suit, My son wore it to prom.
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Reviewed in the United States on April 29, 2026
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