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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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4.5 ★★★★★
Based on 21 reviews
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Product Reviews
★★★★★ 4
Well-Made Carbon Cabin FIlter
I love the honeycomb carbon filter on this cabin filter. It looks like it will do an excellent job cleaning the air in my car. It came in a cardboard box and was easy to install and fit my Toyota well. I would have appreciated some arrows on it, like other filters come with, to let me know I was inserting it correctly. I just followed the example of the one already in my car. If you accidentally take that one out first, you may not remember which side goes where. So, my improvement would be arrows on the filter itself.
Otherwise, the airflow seems great, it functions well and seems to be a great quality!
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Reviewed in the United States on November 14, 2025
★★★★★ 5
Use activated carbon to clean the air naturally!
This is a serious air filter with activated carbon inside of it and I'm here for it. I use little activated carbon bags in my car already as an air freshener because I hate those fake chemical scents from the little trees or from the car wash. The chemicals in those fake scents give me migraines. I absolutely love the idea of putting the active carbon in the cabin air filter itself to clean the air. This fits prefecture in my 2017 toyota prius v. It was just a easy to install as any purge cabin air filter. The hardest part is opening all the parts of the glove box to get the old one out. This filter seems sturdier than my last one and it's a good value for the money.
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Reviewed in the United States on November 11, 2025
★★★★★ 5
Perfect fit for 2016 Corolla!
Fit's my 2016 Corolla perfect. Has a sturdy frame. Other name brand ones I have purchased before don't have the sturdy frame and squish up. This comes with tiny fragrant beads on the top part and has a light fresh scent when the air is on. Very happy so far.
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Reviewed in the United States on November 8, 2025
★★★★★ 5
Best cabin filter I've ever seen
This is the most complex and over-engineered cabin air filter I have ever seen, and I mean that in a good way! The overall shell is a hard plastic, and inside it are two basic parts; the filter (HEPA quality) and the loose fill activated charcoal. The filter media appears to be operating room quality, and if I were to guess, I would say it's like a MERV 14 or greater. The pleats are plentiful, and the spacing of the pleats are perfect, as well as the attachment of the media to the filter frame. The charcoal is a loose style fill, and it fits between the back of the filter media and the filter screen on the very back. This must have 5 times the amount of charcoal compared to the charcoal infused media you can get from many suppliers. The frame fits perfect in my 4runner, and it is hard plastic with good gasket material to further help the effectiveness of the media.
All in all, this is an incredibly engineered filter, and if you're the type that wants nothing but the best for your vehicle, you've come to the right place by choosing this one. It is pricey, at $20 for one filter at the time of writing this review, but this is a textbook case of "you get what you pay for." I am going to start only buying these from now on, as they are the best quality and construction filter I have seen, and I highly recommend this filter!
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Reviewed in the United States on November 3, 2025
★★★★★ 4
A Simple, Affordable Cabin Filter That Does the Job
I have a mechanic I really trust, which is no small thing these days. The trade off is that he charges a little more for routine services like oil changes, and those visits always include a full vehicle inspection. To manage the cost, I make a point of handling the easy maintenance items myself so he does not feel the need to replace them with higher priced OEM parts. Cabin air filters fall squarely into that category. It takes me about two minutes to replace one, and doing it myself typically saves around fifty dollars. That was the motivation behind picking up this filter for my Toyota Sienna.
Right out of the package it fit perfectly. The size is exact, which is important since cabin filters need a snug frame to work properly. There is a clear airflow direction arrow printed on the side, which is one of the two key things I always look for. Without that, it becomes guesswork and you risk installing the filter backward. With the arrow in place, installation is a quick slide and close operation.
Functionally, cabin filters are hard to evaluate in real time. You do not get a dramatic change the moment you swap them out. What I rely on instead is whether they fit correctly, whether they load evenly over time and whether they avoid shedding material as they age. Based on construction and feel, this one meets those expectations. It is pleated well and the frame holds its shape. All signs point to it filtering properly, which is the baseline requirement.
Because cabin filters need changing fairly often, it never makes sense to overspend on OEM versions unless you have a specific airflow sensitivity or unique environmental need. This filter costs far less and performs the key functions just as well for general use. In my Southern California environment, where dust and pollen appear regularly, I prefer to replace filters on the more frequent side, which makes the lower cost even more practical.
The simplicity of the whole process is what I appreciate most. No tools, no struggle, no surprises. The filter pops in cleanly, the compartment closes and the job is done. When paired with the peace of mind of not paying dealership pricing for such a simple part, it becomes an easy product to recommend. A dependable, properly fitting cabin filter at a very reasonable price. Four stars.
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Reviewed in the United States on December 4, 2025