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Description
Human TGM3 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: Gently wash adherent cells with pre-chilled PBS, then trypsinize and collect the cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash the collected cells three times with pre-chilled 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 freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Collect the supernatant for analysis, or store at -20°C or -80°C, but avoid repeated freezing and thawing. 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 transglutaminase 3, epidermal (TGM3) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB converts 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 transglutaminase 3, epidermal (TGM3) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Transglutaminase 3, Epidermal ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Protein-glutamine gamma-glutamyltransferase E, also known as epidermal transglutaminase, is an enzyme encoded by the TGM3 gene. Transglutaminases catalyze the cross-linking of proteins via epsilon-gamma glutamyl-lysine isopeptide bonds. Although the primary structure of transglutaminases is not conserved, they all share a common amino acid sequence in their active sites, and their activity is calcium-dependent. The protein encoded by this gene consists of two polypeptide chains and is activated from a precursor protein by proteolysis. The encoded protein is involved in the later stages of cell envelope formation in the epidermis and hair follicles. | |||||||||||||||||||||||||||||||||
| 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, cell culture supernatants, and other biological fluids |
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4.2 ★★★★★
Based on 16 reviews
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Product Reviews
★★★★★ 4
It does exactly what they say it does! Worth the money
Color: A1-Red
It arrived quickly. It’s a fun toy! It’s the right size for cats. Definitely worth the money. It is exactly as described. So far, the quality seems very good and it seems durable. The instructions are on the side of the box, which makes learning how to operate it easy. I love it that it’s red, because my kitty seems to be attracted to red. My kitty is very interested in it, yet isn’t quite sure what to think of it. I love that it changes directions on its own so that it doesn’t get stuck or lost under furniture, which is typical for a ball that cats play with. It seems to only go two, three or four feet in one direction and then it changes directions, which is both good and bad. I just wish it would go farther in one direction before changing directions so that my kitty could chase it farther, as we have plenty of space in our house for her to run. But that certainly could be a plus for other cats, enjoying not knowing where the ball is going to go next and really keep them on their toes.
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Reviewed in the United States on January 20, 2026
★★★★★ 5
Cat loves this
Color: A1-Red
This toy is great for keeping pets entertained. The ball moves quickly and the tail makes it even more fun for cats to chase. It’s rechargeable and easy to use, and my pet stayed interested much longer than with regular toys.
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Reviewed in the United States on March 15, 2026
★★★★★ 5
A Great Energy-Burner for High-Drive Pups!
As the owner of a high-energy dog, finding something that keeps them occupied for more than five minutes is usually a massive challenge. The LDZ&CHX Smart Interactive Dog Toy has been an absolute lifesaver for our daily routine.
What I Love About It:
True Boredom Buster: The moving and rolling action is erratic enough to mimic prey, which immediately hooks my dog’s attention. It does a fantastic job of keeping them busy and mentally engaged when I'm tied up with work.
Smart Design: The inner ball provides the movement, while the outer blue cage-style sleeve makes it incredibly easy for dogs to grab, carry, and chase without getting their jaws stuck.
Eco-Friendly & Wallet-Friendly: Thank goodness it's rechargeable. I am so tired of buying endless packs of AA batteries for moving pet toys, so being able to just plug this in is a massive plus.
Things to Keep in Mind:
The "Bite-Resistant" Sleeve: While the outer sleeve is durable, I wouldn't leave a notorious heavy chewer unsupervised with it for hours. It works beautifully as an interactive chase toy, but it shouldn't be treated as a pure, indestructible chew bone.
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Reviewed in the United States on June 11, 2026
★★★★★ 4
Great toy for dogs, cats, or even children!
I got this for my dog, but she was completely uninterested. (I'm sure there are tons of dogs who would love it, mine just happens to not.) But a friend came over with her toddler son, and he absolutely loves it, so I gave it to him. It's a pretty fun little ball and vibrates strongly enough to make itself roll around in random patterns on the floor. It also lights up as it does so, and looks pretty cool!
The build quality seems to be pretty good, the outer rubber cover is pretty durable. Though some dogs will probably make quick work of tearing it up, as it has a large opening for the ball to come out.
And the USB-C charging is a nice feature as well, but you have to remove the ball from it's sleeve and unscrew it to charge it.
As for value, I think it's not a bad deal overall, and would recommend it for small/medium dogs, cats or children! It's well worth the price for the amount of entertainment it will provide!
As always if I encounter any other issues or think of anything else to add I'll update this review!
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Reviewed in the United States on June 12, 2026
★★★★★ 5
Puppy-Approved Entertainment
Oh my gosh, my puppy absolutely loves this toy. Out of all the toys we’ve brought home recently, this one may have become her favorite.
She chases it around the room, pounces on it, carries it around, and then does the funniest thing—she’ll bring it back to her bed and just hold it in her mouth while it vibrates. I’m not sure if she’s soothing her teething gums or if she simply enjoys the sensation, but she seems completely fascinated by it.
It’s honestly pretty entertaining to watch. The unpredictable movement keeps her engaged much longer than many of her other toys, and it does a great job encouraging active play. For a puppy with endless energy, that’s a big win.
The outer sleeve has held up well so far, and I appreciate that the toy is rechargeable instead of constantly needing replacement batteries.
If you’re looking for a toy that can keep a curious puppy busy and provide some laughs for the humans watching, this one has been a hit in our house. It’s getting a lot more playtime than most of the toys in her basket.
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Reviewed in the United States on June 8, 2026
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