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Description
Human Trx 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. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend 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 freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. 5. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 6. 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 thioredoxin (Trx) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of thioredoxin (Trx) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Thioredoxin ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Thioredoxin (Trx) is a 12 kDa oxidoreductase protein encoded by the TXN and TXN2 genes in humans. It also possesses a unique tertiary structure, termed the thioredoxin fold. Thioredoxin is maintained in a reduced state by the flavoenzyme thioredoxin reductase in an NADPH-dependent reaction and also serves as an electron donor for peroxidases and ribonucleic acid reductases. The related glutaredoxin shares many of the functions of thioredoxin, but its reduction is performed by glutathione rather than a specific reductase. | |||||||||||||||||||||||||||||||||
| 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 | 0.15-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.1 ★★★★★
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Product Reviews
★★★★★ 5
Nice recommended
Color: 01# Golden Amber, Color: 01# Golden Amber
Very smooth lotion makes your skin feel dewy. Very light smell of coconut. And protect you from UVs rays. It really glows
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 11, 2026
★★★★★ 3
Sticky, I’d hate it all over my body. Shimmer is light but glittery.
Color: 02# Rose Gold
Very sticky formula, wore it over regular sunscreen bf I didn’t want to feel that sticky so I only added it to certain spots like my color bones. I’d probably skip next time and just use a highlight.
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Reviewed in the United States on May 30, 2026
★★★★★ 5
Absolutely love and fully worth it
Color: 02# Rose Gold, Color: 02# Rose Gold
I love this product now you can see a little bit of shimmer on my skin, but it’s hard because I’m pale. I will update this with a picture when I get a Tan.. but it smells absolutely delightful. I’m glad I’ve purchased this over the high priced fenty body lava .. and I love it because of the SPF in it . I’m definitely going to purchase the other shades.. it smells like you are on the beach.
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Reviewed in the United States on May 11, 2026
★★★★★ 5
Best toy for Fetch, Inside and outside. I have multiple of them!
Color: Colors May Vary, Size: 5.5 Inch
Size: LARGE
This is hands-down my favorite toy for playing fetch/exercising my dog. I've already bought 2 of these, and I am buying a third because the first one is being rotated out (it got baked by the sun after 3 years outside haha).
My Story:
My dog LOVES to play. All the time. Inside and outside (well duh, all dogs do). But this toy allows me to play inside while not worrying too much about breaking things. My dog has grown out of his "chew to DESTROY it!" phase, so he is able to keep his toys for a long time.
I hate touching toys after they've been slobbered all over, it keeps me from playing with him! There are so many toys that become slippery or slimy after just one throw! But THIS is a great toy because you are able to pick it up with one finger, and just fling it really easily, without getting your hands all gross.
In fact, when i'm playing outside with my dog, I actually KICK these (large or jumbo size only) instead of throwing them. They are perfect for that! and I can get a good kick and consistent placement every time with this thing. And then, I don't have to touch it with my hands AT ALL!
Reasons I love it:
- Non-destructive Indoor Play: I like that this thing keeps its shape, but when it hits things, it just doinks off really lightly and doesn't cause any marks or damage to walls, windows, etc. Note: my house is also not full of knick-knacks or delicate items placed all around that are in harm's way. I'm sure it could knock down some little glass vases if it made contact.
The point is, if you place your throws well, you are very unlikely to damage anything in your house (besides your wood floors from your dog's nails sliding all around haha).
- Quiet: When you kick/throw this toy, and then it lands on the ground/hardwood, it doesn't sound like a bomb went off in your house. It lands softly and lightly, as if you tossed a half-empty toilet paper roll. I have hardwood at my house, so when I play with other toys, they land on the floor and go BOOM as if you just dropped a 20lb weight! If someone else is playing with my dog with other toys and i'm in the other room, it nearly scares the pants off me. I run in and say, "WHAT THE HELL IS GOING ON --oh, you're just playing! so cute. Sorry, continue on."
- Slobber-Free: You don't have to touch the slobbery part in order to throw it or kick it to your dog! Very easily kick-able; it's my preferred method of playing fetch (least energy for me, although I should be the one running, ha).
- Easy to catch: Super easy for my dog to catch. He should be a wide receiver cuz he catches things that, when I kick it, while it's in the air i'm thinking, "AW CRAP, that's way too far..... OH WAIT, he got it?!?! noiiice!" Makes an amateur "quarterback" proud lol.
- Price: It's not $20 per ball! Some toys are $20 and they make you REALLY have to think ... "Does he REALLY need this toy....??" the answer is always NO, (because he has a million toys that are perfectly good). And then I waver so long on the purchase that I forget it even exists!
- Longevity: If your dog is past the "MUST.DESTROY.ALL.THINGS!" stage, then it will last a long time. After 3 years, I'm finally replacing/rotating-out the first one I bought 3 years ago. It has been in the sun, rain, winter, mud; everything. I could still use it; it's totally usable! I was just like, "Eh, why not get a new one?" Honestly, you can never have too many of these. seriously.
- Light, Easily Portable: I always pack these in my dog bag. They are light, and they can smoosh into the bag to a smaller size if my bag is full. No problem.
- Least-annoying toy that your dog brings to you: My dog likes to HINT that he wants to play by bringing his toys and setting them in my lap (wherever i am). With most of his other toys, my reaction is, "GROSS! it's already so slimy!" and I flick it off of me. This Hole-Y Roller is light and not slimy so you can quickly fling it for your dog without getting side-tracked from whatever “unimportant human activity” you’re doing.
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Reviewed in the United States on September 12, 2018
★★★★★ 5
Fun with a creative twist
Color: Colors May Vary, Size: 5.5 Inch
This would be a toy to get creative with. The quality is there. It’s a nice size it rolls so it’s great as just a bowl that’s slightly rubbery silicone like that the dog can play with and chew a bit. If you have a strong tour, and you believe that they chew everything that you give them eventually they would probably chew it open. Our puppy likes it plays with that as a typical ball. It’s perfect because he can grab it in his mouth and walk around with it lightly and easily. We do take a kitchen towel. Roll it with treats and shove it inside and he’ll have to try and retrieve that that towel with the treats so it kind of gives us a few minutes of peacefulness. You can create so many different fun games with it because you have access to place things inside you know the holes are slightly big so the items inside would have to be of a good size. We have put his his large wishbones inside to allow for him not to have to hold the bone and it gives him some sort of structure as well as if it rolls away, he finds it to be a game so that’s nice. It’s made well. It’s good quality. I like how it’s blue so the dog could see the color and it’s easy to store cause it’s not huge.
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Reviewed in the United States on January 11, 2026