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Description
Mouse PEMT 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 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 capture antibody against phosphatidylethanolamine N-methyltransferase (PEMT). 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 phosphatidylethanolamine N-methyltransferase (PEMT) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Phosphatidylethanolamine N-methyltransferase ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Phosphatidylethanolamine-N-methyltransferase (PEMT) is a transferase (EC 2.1.1.17) that converts phosphatidylethanolamine (PE) to phosphatidylcholine (PC) in the liver. In the CDP-choline pathway, choline obtained through the diet or metabolism of choline-containing lipids is converted to PC, accounting for approximately 70% of PC biosynthesis in the liver. The PEMT pathway has been shown to play a key evolutionary role in providing PC during starvation. Furthermore, PC produced by PEMT plays a wide range of physiological roles, including choline synthesis, hepatocyte membrane structure, bile secretion, and very low-density lipoprotein (VLDL) secretion. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, cell culture supernatants, and other biological fluids |
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4.4 ★★★★★
Based on 2159 reviews
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Product Reviews
★★★★★ 5
King has met his match.
Color: Large - Billy the Beast Monster
I received my dog's toy this evening when I place the toy on the ground my dog King went crazy. He is a one-year-old Pitbull and love to tackle any toy he believes he can't take down, but Mr King has met his match with this toy. He was using all of his bite force to demolish the toy but the toy actually has worn him out. He ran around my yard for about 30 minutes with the toy in his mouth producing the squeaking sound from the toy as well. Hold up the toy is totally filthy with dirt almost covering the entire green and yellow belly of the toy. I have brought king mini toys in the last couple of months and this is the best toy that holds up to his energy level. I tried to retrieve the toy and place it in his dog house because it is far beyond his bedtime, but he wanted to play catch me if You can with my new toy. This is a great toy for those big energetic dogs that love to take their aggression out on maybe some furniture or perhaps that favorite shoe you do not want them to chew on. I am totally sold on this toy I brought from the seller who placed the items on the Amazon site. I plan on purchasing a another toy from the buyer to keep King energy at a low level if I can. I just checked on my dog and he actually has taken the toy with him inside of his dog house and now the toy is snugged under his left paw. I'm one of those smart owners I'm not going to even try to take it from him tonight but hopefully in the next week I can sneak it from him and it may be perhaps wash it. I did this item AA Plus
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Reviewed in the United States on January 31, 2026
★★★★★ 3
Cute, but not Maligator-proof!
Color: Small - Towering Timmy Monster, Color: Small - Towering Timmy Monster
Fun toy while it lasts. My Belgian Malinois made quick work of this little guy but still enjoys playing with its carcass. Bite Force* but not taking into account the bite force of a Mali-gator!
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Reviewed in the United States on May 7, 2026
★★★★★ 5
Puppy Love
Color: Small - the Goofy Monster, Color: Small - the Goofy Monster
Short backstory.... My daughter moved in with her 3 dogs recently and brought her dogs toys. Over the course of the last 3 months, my dogs have managed to steal every last toy they could find.
fast forward to purchasing this Blue Monster. It came today and was immediately grabbed by one of my dogs. She refused to give it up even for her favorite ball. Ten minutes later another dog had the baby monster and was not giving it up!!!! (think growling) I thought that I would put the monster up for the night to allow for some sleep but it felt like it had been soaked in slobber ( I am sure I saw it drip ;) ). My dogs seems to think this little blue monster is their one true love.
Now for some facts.
1. Baby monster is quite small in comparison to my other Bite Force dog toys. It is not too small for my dogs but it is definitely not a size I would give to my daughters Pit that thinks she is a vacuum.
2. Squeaker lasted about 2 minutes before my dog drool invaded it. Currently no squeaky sounds can be heard...just slightly squishy sounds
3. No holes despite my dogs trying. There has been constant chewing (by Standard Poodles) on this little guy for the last 5 hours and not one little open area. Trust me this is a huge accomplishment. Stuffies last 10 seconds in this house.
If you need a toy that will last longer than most I highly recommend Bite Force. The are designed for tough chewers and take a beating
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Reviewed in the United States on November 11, 2025
★★★★★ 2
My raptor GSP destroyed this dino toy
Color: Medium - Dilophosaurus, Color: Medium - Dilophosaurus
My dog ripped this toy apart faster than the trex did the lawyer in Jurassic Park. 2 hours! Arm was ripped off, stuffing pulled out methodically over the next to days. Now it is a gutless husk. The metor was kinder to the og dinos than my gsp was to this. Sorry Bite Force. Open to product sampling tho!
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Reviewed in the United States on May 19, 2026
★★★★★ 4
My Hero pup is super happy with it
Color: Large - Billy the Beast Monster
I was super excited to order this and had waited some time to do so. I wanted Big Billy The Beast Monster in large Green, he’s an awesome looking guy. My Golden Retriever was all over me while I was up packaging Him and his tail was in helicopter mode. He was so excited to have it and carried it around the house everywhere we went, he’s laying on me with his head in my lap and Billy’s right next to him. Reason for buying it? Well he loves plushies but there are not many available out there that can stand up to this 80 pound chew monster so I have to keep him on harder stuff, 2 of which are the cheapest toy skull and crosses in orange and blue from a cheap outlet site but he loves them. Orange was first then when we pulled out the blue one he had a aha moment. Now with Billy he was on a whole new level of excitement, I’ve never seen him this excited over a toy, he can’t stop carrying it with him everywhere and showing it off to the other pups I have, there’s no growling or resource guarding but he definitely won’t want to share and that’s of because I always take away any plushies that he gets to because he is serious about chewing. This being made with Kevlar gave me hopes of longevity but he had the ears and the end of 1 hand chewed off in under 2 hours, am I disappointed…yea a little but he has gotten so much joy out of it that it’s made me really happy to have bought it anyway, see he’s my Service Dog so he spends a lot of time being so serious and working that he needs any outlet for spending energy possible and he’s so happy with it that it makes me happy. As I’m writing this he’s asleep with his head in my lap and Billy right next to him looking very tired and happy and that my friends is worth all the beans in the world.
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Reviewed in the United States on May 4, 2026
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