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Description
Rat TPO ELISA KitProduct Specification Usage Required experimental equipment: 1. Microplate reader (450nm) 2. High precision pipettes and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and
Product Specification
| Usage | Required experimental equipment: 1. Microplate reader (450nm) 2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37°C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL 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 collect the supernatant for analysis. Cell Lysis Buffer: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended 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 freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. 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 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 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 thyroid peroxidase (TPO) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of thyroid peroxidase (TPO) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Thyroid Peroxidase ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Thyroid peroxidase (TPO), also known as iodide peroxidase, is an enzyme primarily expressed in the thyroid gland, where it is secreted as a colloid. Thyroid peroxidase oxidizes iodide ions to form iodine atoms, which then attach to tyrosine residues on thyroglobulin to produce thyroxine (T4) or triiodothyronine (T3), the thyroid hormones. Thyroid peroxidase is encoded by the TPO gene. It is a common autoantibody epitope in autoimmune thyroid diseases, known as anti-thyroid peroxidase antibodies (anti-TPO antibodies). | |||||||||||||||||||||||||||||||||
| 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 | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.8 ★★★★★
Based on 24 reviews
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Product Reviews
★★★★★ 5
Mental stimulation for dogs and fun to watch!
Color: Turn Around, Size: Level 2, Color: Turn Around, Size: Level 2
Perfect mental stimulation for dogs like that like a challenge! My very smart dog figured it out in a few seconds so I use treats that are only a little smaller than the holes to make it more challenging. She doesn’t play with toys like my other dogs so I do make an effort to offer her other outlets to get energy out. I have another dog that finds it challenging, but enjoys it, so I use smaller treats for her to make it easier. I have another dog that has a low tolerance for challenges and walks away from it or tries to chew through it- so I think the level 2 rating is fair as every dog is different! My dogs are 12 lb for reference.
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Reviewed in the United States on May 2, 2026
★★★★★ 5
Great tool!
Color: Flip Board, Size: Level 2
Love this interactive toy! I purchased this for our 4 month old husky, looking for something to keep her entertained, and I was not let down! In the beginning, it took her a few good minutes to figure it out, and about a week later, she gets it done in about 2 minutes. If you've never had a husky, let me tell you, these dogs are very inquisitive! But due to her being involved with this, our older pit is now getting in on the fun! This is huge because our pit isn't amused by much! Overall, it's durable enough to withstand puppy chewing, and being carried around. It's also super easy to wipe clean as it's some sort of durable, perhaps reinforced layers of plastic. Even though the puppy figured it out, it still seems to excite her, and considering it keeps both dogs entertained and active mentally, I would absolutely recommend this to any dog owner!
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Reviewed in the United States on October 17, 2025
★★★★★ 4
Trixie Flip Board Dog Enrichment Toy – Great Starter Puzzle, Best for Beginner Dogs
Color: Flip Board, Size: Level 2
I’ve been using this with a high-energy, working-breed dog, and it’s a solid beginner-level enrichment toy—especially for dogs new to puzzle feeders.
What works well:
• The design is simple and intuitive. My dog learned the flipping lids and sliding compartments quickly without frustration.
• It’s great for short mental stimulation sessions or slowing down treat time.
• The non-slip base helps keep it in place on hard floors.
Things to consider:
• This is a Level 1 puzzle, so once your dog understands it, they can solve it quickly.
• The compartments are on the smaller side, so it works better with small treats than full meals.
• Best used with supervision if your dog tends to paw or chew at toys.
Best use case:
Ideal for puppies, beginner dogs, or as part of a rotation of enrichment toys. More advanced dogs will likely need something more challenging to stay engaged.
Bottom line:
A well-designed starter puzzle that’s easy for dogs to learn and use. Not the most challenging option long-term, but a good addition to an enrichment routine.
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Reviewed in the United States on March 31, 2026
★★★★★ 5
My rabbit loves playing with this, and of course getting treats!
Color: Turn Around, Size: Level 2
My bunny absolutely loves the TRIXIE Activity Around Puzzle Feeder! It keeps her engaged and mentally stimulated for long periods, making it a great source of fun not just for rabbits, but for cats and small dogs too. The design is intuitive and easy to clean, which I really appreciate—it takes just a quick rinse or wipe-down after each use. Despite regular use, the toy has proven to be very durable and holds up well to enthusiastic play. One of the standout features is that it’s surprisingly resistant to chewing; even with my bunny’s constant nibbling, there’s barely any visible wear. The compartments are just the right size for small treats, encouraging pets to think and interact with it. I love that it adds enrichment to my pet’s routine without creating extra work for me. Overall, it’s a well-made, multi-species puzzle toy that brings a lot of joy to curious pets. It is very quiet during use and doesn't make a noise.
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Reviewed in the United States on June 29, 2025
★★★★★ 3
He was confused, I was amused!
Color: Turn Around, Size: Level 2
This toy was quite well made, the materials are solid and once assembled, its very sturdy! Super easy to put together too. It comes with two kinds of lids for the test tubes, three little holes (intermediate/ difficult) and one big hole (beginner/easy). So the actual toy was really nice.
There was only one problem...for some inexplicable reason...my dog (3 yr/ 20lb/ shorkie) was afraid of it! He's used lots of puzzle / game toys in the past, but for some reason this one weirded him out.
No matter what I did, I couldn't get him to even try moving the test tubes. The farthest I got was having him eat a treat off the edge of a test tube once or twice (hence the video, there's a piece of chicken sitting on the top). Mostly he would just growl at the toy or eye it suspiciously, and then run away. I would describe his overall attitude as agitated skepticism with a hint of annoyance...specifically toward me! (As if saying "what the heck is this thing??")
I will say the customer service was pretty good. And I definitely got some personal entertainment watching my dog run around and growl at the toy that was just sitting there. (:
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Reviewed in the United States on April 27, 2025