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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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4.1 ★★★★★
Based on 13 reviews
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Product Reviews
★★★★★ 5
Great for crazy pups!
Size: 8 Inch Diameter, Color: Ocean Blue
Tough and fun! This is a great toy to keep in the yard for play. I have a heavy chewer. He can get a hold of it and carry it around but it doesn't pop. It is soft enough but also strong. My dog loves to herd it around. He loves it!
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Reviewed in the United States on May 19, 2026
★★★★★ 4
Durable, bouncy fetch toy that dogs usually love
Size: 8 Inch Diameter, Color: Ocean Blue
Bouncing Dog Toy is generally very well received, especially for dogs that enjoy fetch and active play. It is made from a flexible, puncture resistant material that can handle a lot of biting and chewing without immediately popping or losing shape. Many owners like that it has a good bounce and is easy for dogs to grip, even when wet or slobbery. It is also lightweight enough for kicking, tossing, or rolling, which makes it great for interactive play sessions.
A big highlight is that it floats, so it works well for water play as well as backyard or park fetch. The 8 inch size is especially good for medium to large dogs because it is big enough to avoid accidental swallowing but still easy for them to carry and chase. Most complaints are pretty typical for any dog toy of this type, meaning very strong chewers can eventually damage it, and some dogs may puncture it over time, but even then it often keeps its shape and continues to be usable
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Reviewed in the United States on May 4, 2026
★★★★★ 5
Soccer ball just for dogs
Size: 6 Inch Diameter, Color: Ocean Blue
I could say "soccer", and our shoodle would wag her tail ready to play soccer with a tennis ball so I tried out this real soccer ball with her. Mostly, she just wants to wait for me to kick it, playing defense, so she can chase it like the tennis ball. After that she wants to usually bite it or roll over on top of it as a way to secure the ball, but sometimes she will have fun pushing it around with her nose or paws. She still prefers the frisbee for play time outside since the frisbee is good for catching and for playing tug-o-war, but this is a good alternative that always gets her wagging with excitement.
I wanted to get a regular 6" soccer ball for our shoodle, but this was a good choice for dogs for the durability and safety of the rubber material. The hexagons give the dog some grip to pick it up. The rubber is soft enough to grip and expel air when they do figure out how to grip it. The ball is designed to suck back in air when the dog releases the ball.
- Durable rubber material ideal for dogs
- Dog can squeeze out air to grip it
- Ball fills back up with air after air is squeezed out
- Can make more noise than a leather soccer ball
- Noise is still tolerable when used indoors
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Reviewed in the United States on April 29, 2026
★★★★★ 5
Good for aggressive cheers. Reinflates everytime even when punctured
Size: 6 Inch Diameter, Color: Apple Green
I have an aggressive cheer that will tear apart anything. I bought this a few months ago and it is covered in puncture holes but it reinflates everytime. Best dog toy ever she loves it. I would buy a dozen of these. The medium size is perfect for my mini bull terrier. Big enough to not get completely in her mouth but small enough that she can still grip it to carry it around
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Reviewed in the United States on April 21, 2026
★★★★★ 5
Dog approved for fun, human approved for durability for the price.
Size: 8 Inch Diameter, Color: Orange
This has been my dogs favorite outdoor toy for years . I love these because they are durable and won't deflate when bitten or when carried around in their mouth.... no matter how many holes the dogs put in them. We leave these outside year round even in Minnesota winters. It is strong enough to hold up yet soft enough to be safe on their teeth. Now I do want to say although it is really durable it is not indescribable. IF you have a heavy chewer or a dog that loves to destroy toys, they would need supervision with this I think. My dogs don't try to chew this up but they do wear out over time. We replace it every 2 to 3 years on average. Definitely dog approved and recommended in my home, and human approved for cost and durability. I hope your dogs love it as much as mine do.
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Reviewed in the United States on May 6, 2026