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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 23 reviews
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Product Reviews
★★★★★ 5
Must for subwoofers. True 2mm spec. High quality.
Model: 36 SqFt(3.4 SqM), Model: 36 SqFt(3.4 SqM)
Let me start of by saying WOW! This made a huge difference in the SQ of my L7S pushing 930w rms. Before I had major rattles in my 2020 Honda Civic SI. Taking the time to do this to my trunk and Lid made me realize just how loud my subwoofer actually is. It literally eliminated the rattles throughout my whole car. I still plan on doing this to my doors and rear deck but once I upgrade the speakers. I still have half of the box just doing the trunk and lid so 1 box of 36sf will be enough for most of the areas that need to get done. You may need a little more overall. This is true 2mm spec. Very sticky and easy to apply. It just take time to finish the work correctly and roll it down. It is not as heavy as I thought it was going to be. The size of the sheets are perfect for what you need to do. The value is on point for the quality. It does not rip very easily but you can cut it to size without any problems. Again, This is a must buy for anyone who is pushing 1000w rms or more.
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Reviewed in the United States on November 29, 2025
★★★★★ 5
Quality product you
Model: 18 SqFt(1.7 SqM), Model: 18 SqFt(1.7 SqM)
Summary: Great quality product, priced right. Cuts and installs easy-follow the instructions people!! PRO TIP-you don’t need to cover every square inch. OVERKILL!! Think about it-when a bell rings you could put your fingers on the edge and silence it right? Same rules apply here. A strip or square will isolate the vibration and notice when placed correctly— knock on the surface and see what vibrates or what part makes the loudest drowning and that’s a good clue of where you should put a piece of this…unless you’re trying to use it for temperature insulation no need to go ape on it. If you look at the bottom of a stainless steel sink, it’s got a stripe or a small sticker of insulation that keeps it from rattling. You can do that here. What’s important to note is the instruction specifically say use a roller to roll the ribs out until it’s smooth that way you’ve got maximum adhesion. I see a lot of pictures and videos from people that finished their install and didn’t follow this simple instruction. It makes sure that it’s stuck down before you put all your paneling and carpet back in and it’s a small step with a cheap wooden roller it’s easy. I’ll say the installation with this product is a breeze. You can use sharp scissors or a blade. It does get gummy after a while, and you may need to clean off your blade or your scissors, but it cuts very well and the ribs make it easy to size in space the thickness is fantastic and the backing paper comes off quite easily. I’ve done lots of installations with Graphics emblems you name it and this product is very easy to use and represents good value.
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Reviewed in the United States on March 30, 2026
★★★★★ 5
Good stuff !
Model: 36 SqFt(3.4 SqM), Model: 36 SqFt(3.4 SqM)
This is an excellent product. Easily moldable, cuttable with good scissors or razor knife, very sticky, works great.
This pack was enough to do all 4 doors on a crew max truck, and I went wild on the fronts. I used 9 peices on each rear door which was just enough, and the rest on the fronts. I doubled up layers and even used some on the back of the door panels. If you follow the instructions and dont go wild you might even have extra left over.
It works just like you would imagine. Without doing the entire cab there will still be road noise but this helped noise, better audio, no buzzing door panels, terrific improvement.
It is kind of heavy, so extra weight is added to truck, but that really isnt a bad thing. It is noticable but in a good way.
Get it. Use it.
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Reviewed in the United States on March 20, 2026
★★★★★ 4
Good product
Model: 36 SqFt(3.4 SqM)
Easy to roll out. Works great
Why did you pick this product vs others?:
Fair price for the amount of coverage, good quality
Works well:
Easy to install, rolls out great. Forms to contours well
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Reviewed in the United States on July 13, 2025
★★★★★ 5
Great product
Model: 36 SqFt(3.4 SqM)
I used this on the doors of a Toyota Tacoma, worked as it described. Reduced the road noise significantly.. Need to buy another box for the floor and roof of the truck..
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Reviewed in the United States on May 15, 2026