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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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4.3 ★★★★★
Based on 1561 reviews
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Product Reviews
★★★★★ 5
Love this ball!
Size: Large (Pack of 1), Color: Multi
My doberman and her dog friends love this large high bounce rubber fetch ball! She loves to chew it endlessly between throws and her Rottweiler buddy loves to do the same because it has a hole through it. I definitely recommend the large 3 inch ball for larger dogs.
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Reviewed in the United States on November 15, 2025
★★★★★ 4
The orange ones are perfect!
Size: Medium (Pack of 2), Color: Assorted
I wish I could give 4 1/2 stars but it isn't possible.
The reason why I don't give 5 stars is because no one sells these in a 2 pack of just orange balls. I was able to find blue only but not orange only. I have a pitbull and so does my good friend. We take our dogs out all the time and they absolutely love chasing these balls.... The orange ones to be exact. They will not play with or bring the blue balls back. They are a waste. I had to buy 2 packs to keep the oranges. I'm taking the blue ones to the local dog park and just dropping them off.
Now to the reason these balls deserve 5 stars: They are super easy to throw and they also make a slight whistling noise when you throw them because of the holes. They are real easy to wash due to the material. Also, they are pretty indestructible. The only reason I'm ordering more is due to them being lost. Our dogs have never torn them apart. I'm sure they could if we let them, but we use these for chase, not as chew toys. My friend also tied a rope through one of the balls to throw and retrieve with the rope. He uses this as a fun indoor toy.
These do not float in water but I buy other chuckit balls for that purpose. They have the orange balls with the blue stripe which work great in pools.
The balls are awesome and I keep coming back for more. Hopefully your dogs like the blue balls.
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Reviewed in the United States on December 8, 2015
★★★★★ 5
Worth the money
Size: Medium (Pack of 2), Color: Assorted
I've had several sizes of Chuck-It ball launchers for years but had resisted buying the balls to go with them because I had about 50 tennis balls already that cost only pennies apiece and these were quite a bit more expensive. But, I got tired of the dogs chewing up the tennis balls. I'd start a ball throwing session with a brand new ball and for the first few throws it would sail through the air and the dogs would have to work to retrieve it. However on the way back, their jaws would work it, chomp-chomp-chomp. And when they returned to me, they'd want to stand there and chomp some more, despite my command to "Drop it!" Within 30 minutes, the brand new ball would have a hole in it and then instead of sailing several football fields through the air, then bouncing over a couple of trees, I'd throw it and it would piddle unenthusiastically to the end of the driveway before falling to the ground with a sulky thud, not even bothering to bounce.
So - I finally grew tired of going through a ball every time I play with the dogs - which is everyday - and ordered these chuck-it balls. They are the same size as a tennis ball but made of a rubber-like material (not silicone) and after several months of use, have no wear and tear on them at all. The dogs can exercise their gums on them all the way back to me - throw after throw - and the ball still flies the same distance each time, and bounces satisfyingly upon contact with the ground.
The description says "colors may vary" but the first packet I ordered were blue and orange as pictured. Unfortunately my dog lost the orange one the first time we used it. She got thirsty and ran down to our pond for a drink, dropped the ball in the pond and it hasn't been seen since. It is dense enough it doesn't float as a tennis ball might, and by now is probably so covered in mud and slime that I wouldn't recognize it if I tripped over it. Lesson learned: we don't throw the ball in the pond pasture any more.
After losing the orange ball, I ordered a second packet of the balls so that I would always have a spare. The second packet is also blue and orange. So while colors may vary, in my experience so far, they haven't (which doesn't matter to me or the dogs).
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Reviewed in the United States on March 1, 2015
★★★★★ 5
Super Chewer Friendly!!
Size: Large (Pack of 1), Color: Multi
This ball is perfect for the super Chewer!! It is squishy and durable rubber that stands up to the aggressive chewer. It does not squeak. It has great bounce and is a great toy for fetch. Very cute to watching my pup bring the ball back for another throw. The rubber doesn't stick or have an average powering smell. It smells just like a rubber ball. It is highly functional for a great game of fetch! Highly recommend and very happy with our purchase. It is well worth the price. We will be ordering more.
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Reviewed in the United States on May 20, 2025
★★★★★ 5
Love the way this makes you shimmer and shine. It's absolutely beautiful.
Color: Light Rose Gold Body Glow, Color: Light Rose Gold Body Glow
This stuff is bomb! It is so good. It looks so good on the skin and it doesn't have a bad smell. It smells actually good. I would continue. I will continue to purchase this product. I love it that much!
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Reviewed in the United States on April 12, 2026
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