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Description
Human TKT 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. 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 transketolase (TKT) capture antibody. 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 transketolase (TKT) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Transketolase ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Transketolase, also known as TKT, is an enzyme encoded by the TKT gene. It participates in the pentose phosphate pathway and the Calvin cycle of photosynthesis in all organisms. It catalyzes two important reactions that operate in opposite directions in these pathways. In the first reaction of the nonoxidative pentose phosphate pathway, the cofactor thiamine diphosphate accepts a 2-carbon fragment from a 5-carbon ketose and then transfers this fragment to a 5-carbon aldose to form a 7-carbon ketose. Two carbon atoms are extracted from D-xylulose-5-P to produce the 3-carbon aldose-3-P. In the Calvin cycle, transketolase catalyzes the reverse reaction, converting diheptulose-7-P and glyceraldehyde-3-P into the pentose, the aldose D-ribose-5-P, and the ketose D-xylulose-5-P. The second reaction catalyzed by it in the pentose phosphate pathway involves the same thiamine diphosphate-mediated transfer of a 2-carbon fragment from D-xylulose-5-P to the aldose erythrose-4-phosphate, providing fructose 6-phosphate and glyceraldehyde-3-P. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, tissue homogenates and other biological fluids |
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4.6 ★★★★★
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Product Reviews
★★★★★ 5
A Clear Path Through Estate Planning
Format: Kindle
I went in expecting to feel overwhelmed, but this book flipped that fast. It is a game-changer with so much detail, you'll have a solid understanding of living trusts and how you and your family will benefit. The biggest win for me was finally grasping how to properly move assets into the trust without second guessing every step. It walks you through real scenarios like blended families and uneven inheritances in a way that actually makes sense. I also liked the practical worksheets and ready-to-use documents that keep you moving instead of stalling out. By the end, I felt confident making decisions I had put off for years. It turns a confusing process into something clear, doable, and worth finishing.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 21, 2026
★★★★★ 4
A practical guide that makes a complicated process feel managable
Format: Kindle
I picked up this because estate planning always felt overwhelming, and I wanted something I could actually follow. This guide does a solid job breaking things down into clear steps, especially when explaining how to set up and fund a living trust and what documents are needed along the way. I appreciated the focus on real situations, like blended families, choosing trustees, and avoiding conflicts. The included forms and worksheets made it feel more hands-on, not just theory, which helped me better understand how everything fits together.
At times, the amount of information felt a bit dense, and I had to reread certain sections to fully grasp the legal details. Still, the structure is logical and keeps you moving forward.
Overall, I found it useful and informative. It gave me more confidence around estate planning, even if I would still double-check some steps before finalizing anything.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 21, 2026
★★★★★ 5
A great tool for learning about trust funds.
Format: Kindle
This is an easy-to-read book that guides one through the process of what to do upon death. I went through this recently with my family and wish that I had this book at the time. Coincidentally, I'm meeting with an attorney for exactly this reason (serendipity?). The book clearly defines the common lingo used throughout the process and identifies principal participants, pitfalls, and a general knowledge of how best to execute one of the most important documents of our lives.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 9, 2026
★★★★★ 5
Valuable information in an easy to understand book
Format: Kindle
This book makes the whole process of setting up a living trust feel way less scary than I expected. The step-by-step instructions are written in plain language, so you don't need a law degree to follow along. I really appreciated the ready-to-use forms and worksheets because they save a ton of time and take the guesswork out of getting started. It also covers tricky situations like blended families and second marriages, which a lot of other guides seem to skip over. If you've been putting off protecting your assets because it feels too complicated, this book is a great place to begin.
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Reviewed in the United States on April 24, 2026
★★★★★ 5
Easy Guide
Format: Kindle
A very practical book with easy to understand guides for living trusts and avoiding probate. This made me feel reassured for when I'm no longer here, to make sure my daughter has what she needs.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 21, 2026
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