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Description
Mouse SIRT3 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 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 enzyme conjugate are sequentially added to microwells pre-coated with a Sirtuin 3 (SIRT3) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of Sirtuin 3 (SIRT3) in the sample. The absorbance (OD value) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Sirtuin 3 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Sirtuin 3 (SIRT3), also known as the mitochondrial NAD-dependent deacetylase sirtuin-3, is a protein encoded by the SIRT3 gene. It is a member of the sirtuin protein family and is homologous to the yeast Sir2 protein. It exhibits NAD+-dependent deacetylase activity. Members of the sirtuin family are characterized by a sirtuin core domain and are divided into four classes. The protein encoded by this gene is included in class I of the sirtuin family. Sirtuins have a range of molecular functions and have emerged as important proteins in aging, stress resistance, and metabolic regulation. | |||||||||||||||||||||||||||||||||
| 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, and other biological fluids |
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4.2 ★★★★★
Based on 30 reviews
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Product Reviews
★★★★★ 5
Epson EcoTank ET-2903
Epson ecotank are great replacements for standard printers. The fact that you get to use the ink bottles rather than the rip off ink cartridges that only last a little while is an amazing benefit. They print well and have a good print quality, so much so that we have one for normal printing and one for sublimation printing. I do all my printing wireless and have the rate occurrence that it has connectivity issues, like all printers. It holds a good amount of paper making refills less often Than the cheaper printers. To me it has been perfect for my small home business.
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Reviewed in the United States on June 12, 2026
★★★★★ 5
Goodbye Ink Cartridges
I’ve used cartridge printers for years, and the EcoTank system is honestly the biggest selling point here. Filling the tanks was much easier than I expected thanks to the keyed bottles, and I didn’t make a mess during setup. Print quality is sharp for everyday documents, and color prints come out vibrant enough for school projects, photos, and crafts. The scanner works well, the wireless setup was straightforward, and being able to print directly from a phone is convenient. I also appreciate not having to constantly wonder if I’m about to run out of ink after a few dozen pages. The printer itself is fairly compact for an all-in-one unit and fits nicely on a desk. While it isn’t the fastest printer on the market, the long-term savings on ink and the hassle-free refill system make it a solid choice for a home office, students, or a family that prints regularly.
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Reviewed in the United States on June 7, 2026
★★★★★ 5
WOW! This printer has been VERY impressive thus far *UPDATE*
Color: Black, Style: ET-2800-B, Pattern Name: Printer
My old printer broke (sooner than I would have liked) so I did A LOT of research before deciding to buy this Epson EcoTank printer for home office use. This is my honest review of it this far and I will definitely update my review if I run into any issues.
So far, this printer has been pretty impressive (I've been using it for about a month). The price was pretty average for a printer... but I personally think all printers are overpriced so it was a bit more than I would have liked to spend. It is smaller and more lightweight than I was expecting which is a plus because it takes up less desk space than my previous HP printer.
It is very easy to set up, comes with step-by-step easy instructions and guided instructions through the app. It only took me about 10 minutes (Mostly just waiting for the printer to initialize on its own). It is very easy and surprisingly satisfying to fill the ink tank. I like the idea that this ink might last longer than traditional cartridges and won't have as much plastic waste that went along with my old HP Inkjet.
I have printed plain black and white documents as well as photos. I print all my photos at home, which is most of the printing that I do, so quality was important to me. The photos came out with pretty good quality! No ink smudges or off colors, colors are bright/vivid, and it is easy to modify what paper you're using with automatic prompts on the printer.
Now for the negatives and the reason I'm giving this 4 stars... app to printer connection issues. This was a big issue with my old printer as well. I'm pretty tech savvy and I just have no idea why this is still always an issue for me. There is an extra Epson app that you are supposed to download for printing. Sometimes it can find my printer, sometimes it can't. Sometimes it gives me printer errors or won't let me print documents. And now every time I go to print something it says the paper being used doesn't match the paper settings so I have to bypass that error message (even though it does match in settings and the app, and this happens for both regular paper and photo paper). I've turned everything off and back on again, but the issues with consistent connection and error messages are still an issue.
Usually with some fiddling around, I can get the documents/pictures to print but sometimes it feels like I'm jumping through unnecessary hoops. I'm still working out the kinks so if I find some magic solution I'll update my review. Considering this is an issue with most home printers I've used, it's not a deal breaker.
All in all, if you are looking for a new home printer, I would definitely recommend this one. The quality has been great for me and I am pleased with my purchase (so far).
Hope this helps!
*UPDATE*: I have now printed HUNDREDS of photos, they are all excellent quality, and my ink tanks are still mostly full. This is WAY better than my HP printer as far as ink longevity. Changing my review to 5 stars just based on how long this ink lasts. User interface is still a tad annoying but so worth it regardless.
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Reviewed in the United States on May 10, 2024
★★★★★ 5
Almost a year after purchase and this printer has not let me down!
Color: Black, Style: ET-2800-B, Pattern Name: Printer
I wanted to wait to write a review on this printer to see how it held up over time. Happy to report, after almost a year, it is still working perfectly! I was very nervous about purchasing a printer since it was my first time having to shop for one and almost every printer I looked at had something negative to say about the product. After extensive research, I decided to take a leap with the Epson EcoTank 2800.
I was looking for a printer to use for recreational use. Something with decent print quality and longevity as well as something on a reasonable budget. I normally print full pages of color and have been using it regularly for around a year and I have about half of the ink that came with the printer left. Safe to say, the ink lasts a while. The quality may not be the top of the line quality you can get with a more expensive printer, but for the price and just everyday use, the quality is satisfactory. It was easy to set up and walked you through the instructions to connect to devices and configure it. The screen is a little small but if you can overlook that, it really is a nice printer. I have used regular printer paper, card stock, and sticker paper with this printer and I try to load only a few sheets at a time to avoid a jam but I haven't had any issues with it so far.
Overall, I am extremely happy with my purchase. It can be extremely difficult to find a decent printer and some reviews can be misleading or leave you with questions on if the product is for you. If you are looking for something for printing everyday with decent quality and ink consumption that won't cost you a fortune, this printer is definitely worth taking a chance on!
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Reviewed in the United States on April 24, 2026
★★★★★ 1
My Epson printer is total junk!
Color: White, Style: ET-2800-W, Pattern Name: Printer
I have had many printers over the years. I started with Canon dot matrix printers, and then ink jet printers. In the old days, and ink cartridge contained all the colors, so if red was out, you tossed away the rest of yellow and blue too. They were pricey and wasting ink wasn't something I was OK with. I would pull ink cartridges from dumpsters outside office supplies stores, take them home and draw the ink from them to fill my cartridges with. Then Epson came up with a cartridge for each color and black . That's when I switched to Epson and have used them ever since. I did have an HP in there when I got such a deal I couldn't pass it by, but after several years it died and I switched to Artisan printers. I still have an old 810 that is finicky and I don't want to have to coax it just to print a few pages. I got a WorkForce printer ,used it several times after I got it, and then it sat idle for 5 years or so. I moved recently and decided to set up the WorkForce printer. The new ink cartridges were practically new, but sat for 5 years maybe, so the nozzles and print head needed cleaning, I expected this. It uses mass amounts of ink to do this so then I was out of ink. I ordered new cartridges, and back to cleaning when they were inside. I never got anything to print, plus, the new paper I ordered was as glossy and slippery as photo paper, and the rollers couldn't grip it.
I gave up and ordered this printer as a replacement.
Setup went OK until I put paper in, then the rollers were slipping and I got jam messages, even though nothing was inside. I pulled the paper and let it finish the initial process. Then I put paper in when it wanted to align the print head. It fed paper OK for now, so I guess maybe I didn't get hosed on this Epson as I did buying the WorkForce Epson.
UPDATE
November 1st 2025
This is a frustrating printer!!!!
I'm using WIFI network to connect the printer to my laptop computer. I like to be able to print or scan from anywhere in the house. If you purchase this frustrating Epson printer, you now have a ball and chain around your leg. You can't print a simple document without some kind of error code popping up and requiring that go look at the minuscule little screen and try to figure out what button you must press to get the document printed. If I have to be right next to the printer, I might as well use a wired connection. I am disabled and it's a real inconvenience for me to have to go to the printer and squint at the tiny screen to try to figure out why it's been ten minutes of my time and I still haven't gotten my 1 page document in my hand. High speed printing is not high speed when you can't get a printed page without having to make several trips back and forth to the printer. I set the default paper to the standard plain white letter paper, only to find it wants to scan to photo paper, so it just sits there like a block of cement and nothing gets done. I had an Epson Artisan 810 that I really liked, except for the automatic feeder quit working so you had to stand there and feed each sheet to be scanned by hand. I didn't want a sheet feeder on the printer because they break down and the you're feeding by hand anyway. I just hate printing anything at all now because of this printer. I wasted several hours one day trying to print documents that I downloaded and needed a physical copy to present in person for a project I'm doing. Normally I would send a PDF file, and let someone else bother printing if that was necessary. After years of purchasing Epson printers, this is absolutely the last Epson I have spent my money on. Never ever again. The first clue should have been when the quick start guide suggested using a smart phone to set up the printer. I'm not going to work from my 5 x 7 phone screen when I can instead use a 15 inch laptop screen, and keyboard. It's simply a stupid idea. Sorry, but no more Epson for me ever!!!!
Update:
4/10/26
This is absolutely the most useless and frustrating Epson printer that I have ever ever owned. I cannot see good close up I need reading glasses to read. The digital screen on this printer is minuscule it's like a postage stamp size absolutely worthless. I have to carry a magnifying glass over to the f****** printer so that I can see what that little miniscule screen has to say on it. The most frustrating thing is every time I want to print something which is rare I don't print things every day the printer is not available is the message I always get I go over there by the printer I turn it off I turn it back on, that should clear out any crap that's on it so it's ready to print right? No you would be dead wrong! Instead you get a stupid little mini School screen and wants to be set on copy mode. I don't want to copy anything I don't want to scan anything I just want the damn thing ready to print. Anyway you come back to your your phone and you want to print and a printer still not available. I literally destroyed my phone I was that pissed off. I'm too old to be frustrated like that all the time. At 75 years of age I have had my share of printers over the years starting with the original dot matrix ones. Absolutely none of them were as difficult to use as this stupid printer I got now from Epson. When you want to print something you should be able to send it to the printer and it should print. You should not require a phone app to set the damn thing up! One thing I'll say, it's pretty good on ink because you can't make the damn thing print when you want it to !!!!!!!
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Reviewed in the United States on October 10, 2025