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Description
Human DNM1L 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 Dynamin 1 Like Protein (DNM1L) 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 Dynamin 1 Like Protein (DNM1L) in the sample. The absorbance (OD value) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Dynamin 1 Like Protein ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Dynamin 1-like protein (DNM1L) is a transferase (EC 2.1.1.17) that converts phosphatidylethanolamine (PE) to phosphatidylcholine (PC) in the liver. Choline obtained through the diet or metabolism of choline-containing lipids is converted to PC in the CDP-choline pathway, accounting for approximately 70% of PC biosynthesis in the liver. The DNM1L pathway has been shown to play a key evolutionary role in providing PC during starvation. Furthermore, PC produced by DNM1L 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.8 ★★★★★
Based on 13 reviews
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Product Reviews
★★★★★ 5
Great tool!
Size: 18in, Style: Sport
Ok - so I do not have a dog!!! I purchased this item for a completely different purpose! I saw someone using this to gather eggs!! I often go to my daughter and son-in-law's homestead to carry out the chores when they need to be out of town! Well, I am a bit on the short side (4'11") and some of the hens KNOW that a short person is going to be gathering eggs!!! I simply could not reach all of the eggs in a couple of the nesting boxes! So, after seeing someone else use this ball launcher as an egg retriever, I bought one! I am now able to reach ALL of the eggs - one at a time with no breakage! It was a game changer for me.
I am sure that it works equally well for dog play!!!
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Reviewed in the United States on November 15, 2024
★★★★★ 5
A Fun and Engaging Playtime Essential for Active Dogs
Size: 18in, Style: Sport
The ChuckIt! Sport 18M Dog Ball Launcher has been a game-changer in our playtime routine with our energetic and playful dog. This innovative and well-designed ball launcher has become an essential tool to keep our furry friend entertained and active.
The 18" length of the launcher provides a comfortable grip for both me and my dog during play. It allows for effortless throwing, making fetch games enjoyable for both of us. The launcher's ergonomic design ensures a smooth and efficient ball-throwing experience, saving me from straining my arm during extended play sessions.
The inclusion of a medium-sized ball (2.5") with the launcher is a thoughtful addition. The ball's size and weight are perfect for our medium-sized dog (20-60 pounds), allowing her to easily grab and carry it in her mouth. The ball is also durable and can withstand the enthusiastic chewing of our playful pup.
One of the standout features of the ChuckIt! Sport 18M Dog Ball Launcher is its exceptional throwing distance. It allows us to launch the ball far into the distance, providing ample space for our dog to run and fetch. This extended range adds a fun and challenging element to our playtime, keeping our dog engaged and excited.
The durable construction of the launcher is commendable. It has proven to withstand rigorous use and outdoor play, making it a reliable and long-lasting investment for our dog's entertainment.
Additionally, the launcher's compatibility with other ChuckIt! balls makes it a versatile tool for various play styles and preferences. Whether it's a bouncy rubber ball or a flying disc, the launcher can handle them all.
Moreover, using the ChuckIt! Sport 18M Dog Ball Launcher has strengthened the bond between our family and our dog. It encourages interactive play, providing an opportunity for us to engage and connect with our furry companion while promoting exercise and healthy physical activity.
In conclusion, the ChuckIt! Sport 18M Dog Ball Launcher has been an absolute joy for our family and our playful dog. Its easy-to-use design, impressive throwing distance, and durability make it a must-have for any dog owner with an active and fetch-loving pet. We highly recommend the ChuckIt! Sport 18M Dog Ball Launcher, along with the included medium ball, for a fun-filled and active playtime experience that both you and your dog will love.
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Reviewed in the United States on August 3, 2023
★★★★★ 5
Essential kit for those with fetch dogs
Size: 18in, Style: Sport
Now in our 3rd year, my Brittany and I are down to about 40 fetch tosses a day, some 14,600 tosses a year. The Chuck It launcher wears well, so, figure at least 5 years of use, for 73,000 tosses. That makes for a terrific value!
There are several key benefits of the Chuck It launchers: One is that you can chuck the ball further than you can throw, which allows you to exercise your dog more easily. The second is that you can pick up and launch the ball without touching it, which means NO slime! And, the Chuck It also gives extra reach when you have to retrieve a ball from sticker brambles.
Those are great features for anyone with a fetch dog. If you also happen to have a weak arm or shoulder, the Chuck It launchers provide nice throwing options. I generally throw underhanded. I can't get the full potential range, that way, but, I still get good, long tosses. I curl my second finger above the last bump on the handle, and use a good wrist flick along with arm swing. I've seen others throw side-arm with the Chuck It.
I had originally written a second review for the longer 25M Chuck It. Either I, or Amazon messed up, and it looks like I can have only this one review. So, here are my additional thoughts on the 25M: I bought the 25M so that I could throw the ball even further! The good news is, you really can chuck further, with the longer chucker. The bad news is, those longer throws do require more arm and shoulder strength than when using the 18M. This becomes more obvious when you've done 3 or 4 dozen throws. I would recommend the 25M only for those with the youth and/or strength to get the most out of it. Otherwise, I'd say, go with the 18M.
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Reviewed in the United States on March 11, 2016
★★★★★ 4
Will buy again
Size: 25in, Style: Sport
Fits the bill.
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Reviewed in the United States on May 28, 2025
★★★★★ 5
Must Have for Tiring Out Your Pup
Size: 25in, Style: Sport
The Chuckit! Sport 25M Dog Ball Launcher is an absolute must-have for any dog owner with an active pup! This thing takes fetch to a whole new level—effortless long throws, hands-free pickup (no more slobbery tennis balls!), and a comfortable grip that makes playtime easier and way more fun.
My dog absolutely loves it, and I love that it helps her burn off energy without tiring out my arm. The included medium ball is the perfect size for medium to large dogs, and the durable construction means it's built to last. If you're looking for a simple but game-changing way to upgrade fetch, this is it. Highly recommend!
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Reviewed in the United States on February 13, 2025