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Description
Mouse ACAT1 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 conjugate are sequentially added to microwells pre-coated with an acetyl-CoA acetyltransferase, mitochondrial (ACAT1) 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 acetyl-CoA acetyltransferase, mitochondrial (ACAT1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Acetyl-CoA acetyltransferase, mitochondrial ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Acetyl-CoA acetyltransferase 1 (ACAT1), also known as acetoacetyl-CoA thiolase (ACAT), is an enzyme encoded by the ACAT1 gene. It is an acetyl-CoA C-acetyltransferase. This gene encodes a mitochondrially localized enzyme that catalyzes the reversible formation of acetyl-CoA from two molecules of acetyl-CoA. The ACAT1 enzyme has several unique properties. First, it is activated by potassium ion binding near the CoA binding site and the catalytic site. This binding results in structural changes in the active site loop. Furthermore, the enzyme can use 2-methyl-branched acetoacetyl-CoA as a substrate, making it a unique thiolase. It is regulated at both the transcriptional and translational levels. Enhanced ACAT1 enzyme activity is transcriptionally promoted by leptin, angiotensin II, and insulin in human monocytes/macrophages. Mutations in this gene are associated with a deficiency of the encoded mitochondrial acetoacetyl-CoA thiolase protein, also known as ketothiolase deficiency. Its expression has been associated with the manifestation of prostate cancer. | |||||||||||||||||||||||||||||||||
| 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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4.9 ★★★★★
Based on 30 reviews
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Product Reviews
★★★★★ 5
Excellent product for the price.
Style: ET-4800, Color: White, Set: FAX/ADF/Print/Copy/Scan
I have had many, many printers over the many years at the business and for home. I honestly think this is the best value for what I need. The eco tank is fabulous. This is the first printer I have purchased with the ecotank. So much better than the cartridges. The quality is good and it was easy to setup and put on the network. I did wait a few months before writing this review to truly test it out and write a fair review. I am pleased. I have had Epson products in the past and do appreciate their quality and affordability.
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Reviewed in the United States on June 4, 2026
★★★★★ 1
JUNK, Cheaply built, Feed Scanner fails and falls apart in less than 5 months and only 4 uses.
Style: ET-4800, Color: White, Set: FAX/ADF/Print/Copy/Scan, Style: ET-4800, Color: White, Set: FAX/ADF/Print/Copy/Scan
Absolute piece of Junk!
Built cheaply.
Feed Scanner has never worked properly from my first scan (only did 4) and now falls apart completely, in the roller section. It falls out after it jammed for the 7th time today, this time with a SINGLE piece of paper.
If you have a piece of paper that has a fold in it, forget about it. That will never feed through.
I had 6 sheets I had to get out today, I had to do manually via the flat bed, both sides.
It took 23 minutes to get done. Should not have taken 2.
The last time I used it same issues but less sheets.
The paper is SO TIGHT in the feed section it is hitting BOTH walls.
You could not slip a piece of paper in there if you wanted too, most likely one of the reasons it does not feed correctly.
While on the phone with support (I am right now, 4th call today and 3 emails of pictures they wanted later, the hoops they put you through is ridiculous!)
Of course overseas support and has little to no knowledge of being Tech support, I asked for supervisors only to have my first call disconnect at 1 hour and 20 seconds.
Then the next call was 24 minutes, then another 32 minutes and now this one still ongoing (56 minutes and still going).
Now after 3 hours and 5 minutes, they are "holding" my credit card for 179.99 + tax to get a REFURB unit!
So much for a 5 year Warranty.
Oh they acted like it was out of date on Warranty, it has NOT been in EXISTENCE for 5 years , it came out in 2022.
Did I fail to mention you WILL be dealing with Overseas people who have you repeat the same thing over and over and over and over again. They wonder why I was getting upset...hmmmm .
As they pass you along the ranks you have to repeat EVERYTHING all over again.
As I was on the phone and they were asking me to open WRONG doors to find the serial #, I lifted once again the Jam area and guess what, the parts fell out.
Just PLOP!
Other issues while I am at it, the Picture quality is "OK" nothing special.
I bought it to save ink as HP gouges you to death and destroys your print head if you use a knockoff.
The Ink has been "OK", but I haven't really had it long enough to tell as most everything has been Black and White, so all are still full except the Black (3.4's).
Its a rear feed, I miss the HP draw on that, the print speed again is not fast, just OK. Though never really in a rush with it.
I cant say it enough, this is cheaply built, and my experience with Tech Support was a HORRIFIC.
I never like dealing with Over seas support as it is, but what they wanted pictures of as proof was mind blowing (pictures of the tubes that go to the ink wells? What does that have to do with the issue at hand.)
The panel function is anything but user friendly.
Firmware Updates, if not done, will prevent usage until done.
You have to download and install ALMOST 30 different files, from drivers to firmware updates the list is endless. Never have I had to do that many of ALL printers I have had combined.
Simple ZIP file and let it run on an executable file (EXE)like everyone else. That will take you over 2 hours to get all loaded.
The ink wasn't bad seeing it needs to be primed first to work, I am not sure what will happen when the REFURB comes, will I get ink or just have to re-prime from the cartridges.
The software for scanning is easy to use, though unnecessary, should just scan and auto save, or am I just spoiled by HP?
But to send me a "REFURB" as a replacement on a 5 month old unit with a 5 year warranty is really being low.
They did EVERYTHING they could to get me to NOT follow through in this return, you have ONLY 30 days on Amazon.
You have been warned, I would look elsewhere, the tank issue is a great Idea, but if it doesn't work, what good does it do.
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Reviewed in the United States on December 8, 2025
★★★★★ 5
First Impressions - Complete Review
Style: ET-4800, Color: White, Set: FAX/ADF/Print/Copy/Scan, Style: ET-4800, Color: White, Set: FAX/ADF/Print/Copy/Scan
Printer Review from an Artist.
Printer cost with a 3-year extended warranty - $199.00. Came with four full bottles of Ink. Each bottle was huge, and enough to completely fill up the tanks. In my old Epson printer that used Epson 127 Ink Cartridges a cartridge held 8ml of ink. The cost per cartridge for the XL Black was $31.00 on Amazon. In comparison, a bottle of ink for the tank printer costs $15.00 and holds 65ml. The Epson 127 Ink Cartridge is rated at 175 pages for the XL cartridge and the Tank Printer is rated at 4500 pages per bottle of ink. So, the difference in cost is striking.
Unboxing the printer was simple enough. The printer weighs about half the weight of my old Epson printer. The new printer does not have printer drawers but, back feed only. This actually worked out well for me since I was constantly taking the paper in out and out of the drawers when changing paper size. Also, with this direct paper feed, I figure it will handle my heavier weight card stock paper that I use for my art and greeting cards. To my great surprise, the printer offers two-sided printing. This is really handy when printing my Zines. The printer setup was extremely easy as they recommend a phone app, that walks you through each step with pictures. And it communicated perfectly with the phone for the setup. It takes about 15 minutes to completely set up the printer. I have learned to not rush setup. Take your time, and follow the instructions step by step, Which include filling the tanks.
Features,
* The printer dues Dual-Sided Print - Great for zines.
* It has fax capability, but we do not have a landline, so that will not be used.
* It does copying, with a document feeder or you can lift the feeder and place the document on the glass.
* I mostly use my Epson Precision San copier but it is nice if you wish to make a quick copy of a document.
* Some reviews complained about the very small LED display but I find that by using the app or computer I can control all that I need to do.
Print Alignment. I did this test but the printer heads did not require any alignment.
During the setup process, the program installs the printer driver on my computer. However I found it to be the very basic driver, and I went to the Epson Website and downloaded and installed the latest driver which is very robust and has all the settings that I needed.
The printer's footprint is smaller than my old printer. And my old printer had two paper drawers, though I never used the second drawer.
Negatives.
The output tray is flimsy and seems like it would easily break if bumped when I walked past the printer, so I have to be careful about this.
So far I do not see a way of checking the ink levels from the computer. And they said in the instructions to be sure to check your ink levels. You can see on the front of the printer, where you can see the ink levels. When you purchase more ink, you can attach the bottle and it will not overfill the tanks, and you can then use more ink from the bottle later. So I will purchase a complete set of ink and keep it and as it runs low with about 1/4 left in the tank, I will just fill it up.
So far I am very happy with the investment and now printing and ink cost is not a major factor. And I figure I will do a lot of printing with this printer when the holiday season rolls around for my greeting cards.
As for print quality, I will need to report on this later, though there were no negative reviews of print quality. A test print on regular paper and the colors seemed to be spot on.
One note, I still will continue to use my Canon Pro-300 Wide Printer for my Fine Art Prints, but the new Tank Printer will be my studio workhorse.
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Reviewed in the United States on August 11, 2024
★★★★★ 4
Frustrating to set up and unreliable
Style: ET-4800, Color: White, Set: FAX/ADF/Print/Copy/Scan
I bought the Epson EcoTank ET-4800 hoping for an easy, low-cost printing solution, but it has been nothing but trouble. The wireless setup is overly complicated, and the printer often loses connection to the network, forcing me to restart it or reinstall the software. Printing is slow, even for simple documents, and the paper feeder sometimes pulls multiple sheets at once. While the EcoTank system is supposed to save money on ink, the print quality is inconsistent — colors can be dull, and black text occasionally appears streaky. Scanning and copying are also sluggish. For the price, I expected something much more reliable.
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Reviewed in the United States on August 9, 2025
★★★★★ 5
ET-4800 EZ PZ set up and start up. Prints and scans just fine
Style: ET-4800, Color: White, Set: FAX/ADF/Print/Copy/Scan, Style: ET-4800, Color: White, Set: FAX/ADF/Print/Copy/Scan
An excellent home printer. And why did I purchase such?
I had an Epson WF-520 for nearly ten years. But neglect and lack of use caused the printer head to get gunked up. I cleaned the print heads using a solution of 95% Etoh and glass cleaner (about fifty/fifty), a syringe with a plastic tube attached and LOTS of paper towel strips. It was moderately successful but I was never able to get it back to like new printing.
I looked over several new printers and wanted to stay with Epson because of ease of use (copy, scanning, printing). Seems as tank printers are the way to go these days. So I ordered the ET-4800. Got here in only a couple of days. Setup was pretty easy, just follow the instructions either via the documentation included or online version. I have pinted and scanned a color test page. I used the built in SmartScan feature without having to think about it much. I am quite satisfied with the results.
By the way, there is no automatic two side printing. You actually have to get off your b---, take out the first printing, turn it over and refeed the stack. I was used to doing that with the old WF-520 and never had a problem.
I will now set up a once a week, automatic scheduled printing task. I will just have to make sure there is paper on hand. Lack of use is the killer of these inkjet machines regardless of manufacturer.
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Reviewed in the United States on February 16, 2026
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