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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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4.5 ★★★★★
Based on 20 reviews
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Product Reviews
★★★★★ 5
Amazing!! Redness and itchiness gone after 1 day!
Size: 6.76 Ounce (Pack of 1), Size: 6.76 Ounce (Pack of 1)
So, I've had really bad rashes around my eyes and cheeks for a few years. The doctors can't tell me what it is but just used to prescribe me steroid cream which thinned my skin and gave me eye wrinkles. I read about this in a post on Pinterest and ordered it immediately as I had a recurring rash on my face for 3 weeks that kept coming back. Yes it's $32 but I was DESPERATE. Everything else I tried burned my face, even products with the eczema association seal! I was so excited when this came in the mail. I was tiding myself over with Clinique's Dramatically Different Moisturizing Cream and yes, it hydrated my dry face but it stung on contact a bit, and also made my skin tight and began to peel, not painfully but still gross.
So I finally got a chance to put this xeracalm product on my face last night. I put about a quarter size amount at first, then two extra pea sized amounts under my eyes. Unfortunately through the night my face soaked it all up (my skin is EXTREMELY DRY and it's winter now) but as soon as I woke up and look in the mirror my redness was 90% gone. My face was a little tight but not uncomfortably, and definitely no peeling. It also does not burn on contact which is so great for me.
It's called a balm but it's really more like a heavy thick cream. The top is interesting as you just squeeze it and it pops out and dispenses product to not contaminate it. If you have a problem with the dispenser you have to be an idiot, I just have to be honest lol. It's not rocket science. It serves a very important purpose. Also, I was surprised at the size. $32 seemed pricey but for the amount of product I got it is totally appropriate. I only see myself using this for when my skin gets REALLY DRY and not for regular use though. And a little goes a long way. I'm hoping to see my face go back to normal in a few days. I will definitely update!
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Reviewed in the United States on November 29, 2016
★★★★★ 5
This is SO helpful for Dyshidrotic Dermantitis
Size: 6.76 Ounce (Pack of 1)
I have dyshidrotic dermatitis and this helps me with the dryness and itchy feet. It is pretty greasy but I have to keep socks on so I don't care. It works the best if you use it twice a day but still works well once a day.
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Reviewed in the United States on May 21, 2026
★★★★★ 5
A rich moisturizer that doesn’t feel heavy!
Size: 13.5 Ounce (Pack of 1)
I bought this because the skin on my face is very reactive and has also been very dry and I wanted to try something that was a little more rich than the moisturizer I was using. I liked that this is for face and body so it comes in a bigger size than typical face moisturizers come in. I have been using it for about a week and have noticed a reduction in flakiness and my sensitive skin does not get irritated by it. It absorbs really well and doesn’t feel heavy or make my skin feel sticky. I definitely feel it is worth the price since you get so much in the bottle and it will last a very long time!
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Reviewed in the United States on April 5, 2026
★★★★★ 5
The best eczema cream I’ve used !!!
Size: 6.76 Ounce (Pack of 1)
I will give 10 stars if it were available!!
Used most of the eczema creams out there, including prescribed ointments, but given the fact you can’t avoid using water on your hands throughout the day, the eczema was only getting worse for the past couple of months! I am glad I didn’t give up and found this cream!!!
The second I put this cream on my hands, it immediately moisturized the itching and flaking skin without drying or being greasy unlike other creams I’ve used before. It’s very calming, soothing, and deeply moisturizing! So I had to write this review for those who are suffering the similar symptoms. This may help you as much as it is helping me as I speak!
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Reviewed in the United States on May 5, 2026
★★★★★ 4
Seems to work for damaged barrier (accidental damage as well as eczema and rosacea).
Size: 6.76 Ounce (Pack of 1)
SO I am currently tryign to repair a damaged skin barrier. I have both rosacea and eczema so it's nearly impossible to find a product that makes both conditions happy. I started out using the cream version of this but my face always felt dry with it. I decided to try this since a balm usually helps keep in moisture much better. I am only using a gentle cleanser and this, planning on adding in a toner at some point.
At first I thought I was sent the cream again because this comes out like their cream version but as you rub it to spread, you can tell it has a thicker feel to it. It spreads easily and doesn't tug at my skin. Seems to sink in pretty quickly but it does leave a sticky film until it completely absorbs. I can feel my eyelids stick while waiting. Not sure how long it takes to fully absorb because once it does on my eyelids, I forget. But the stickiness does go away and leaves a soft texture behind. I do find some itching in places I normally get when my skin dislikes an ingredient. But I'm not sure if it's my normal or current damaged barrier.
It helps with the dry feeling and I would say I only feel a slight tightness. But so far, the best moisturizer I have used. And I love that there is NO NIACINAMIDE, NO HYALURONIC ACID, no oat, no shea butter, and no cica/centella asiatica. These ingredients wreak havoc on my skin and everybody is adding this to everything. It's been so hard to find a moisturizer.
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Reviewed in the United States on September 18, 2025