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Description
Human 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 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 utilizes 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) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Sirtuin3 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Sirtuin-3, also known as SIRT3, is a deacetylase protein encoded by the SIRT3 gene. SIRT3 is a soluble protein localized in the mitochondrial matrix and contains a mitochondrial processing peptide at its N-terminus. It is a member of the mammalian sirtuin family of proteins and is homologous to the yeast Sir2 protein. SIRT3 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 classified as the first class of sirtuins. | |||||||||||||||||||||||||||||||||
| 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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4.2 ★★★★★
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Product Reviews
★★★★★ 5
Great product, affordable price
Size: 1 Fl Oz (Pack of 1)
I’ve tried a lot of hyaluronic acid serums over the years, and honestly, this one surprised me. The Ordinary Hyaluronic Acid 2% + B5 leaves my skin feeling soft, hydrated, and plump without feeling greasy or heavy. As someone in my 50s, I really appreciate products that hydrate without making fine lines look worse, and this one layers beautifully under moisturizer and makeup.
What makes it even better is the price. It’s incredibly affordable compared to other high-end hydrating serums that cost 3–4 times as much. You truly get amazing quality for the money.
My skin feels smoother, looks healthier, and stays hydrated throughout the day. I will definitely keep repurchasing this product and highly recommend it for anyone wanting affordable, effective skincare.
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Reviewed in the United States on May 17, 2026
★★★★★ 5
Hydrating, Simple, and Great Everyday Serum
Size: 1 Fl Oz (Pack of 1)
I’m giving The Ordinary Hyaluronic Acid 2% + B5 5 stars because it’s such a good, simple hydration serum.
It makes my skin feel noticeably more hydrated and plump, especially when I use it on slightly damp skin. I also like that it layers easily with my other skincare and doesn’t feel sticky or heavy.
Overall, it’s an affordable, effective product that I can use every day and always feel good about.
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Reviewed in the United States on May 16, 2026
★★★★★ 5
Hydrating Serum That Leaves Skin Smooth and Plump
Size: 1 Fl Oz (Pack of 1)
This serum feels really hydrating and helps my skin look smoother and more refreshed. It absorbs nicely without feeling too heavy, and my skin feels softer and more moisturized after using it consistently.
I also like that it includes ceramides and vitamin B5, which help support the skin barrier while adding hydration. It layers well with other skincare products and works great for both day and night routines. Overall, a great affordable serum for keeping skin hydrated and healthy-looking!
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Reviewed in the United States on May 11, 2026
★★★★★ 5
Affordable and Lightweight Hyaluronic Acid Serum
Size: 1 Fl Oz (Pack of 1), Size: 1 Fl Oz (Pack of 1)
I absolutely love The Ordinary brand and own quite a few of their skincare products, so I was excited to add this hyaluronic acid serum to my routine. I use it after cleansing my face and before applying moisturizer, and it layers really nicely with the rest of my skincare. One of the things I appreciate most about this product is the packaging. The glass bottle feels high-quality, and the dropper makes application easy and convenient.
The serum itself is very lightweight, and a little bit goes a long way, so the bottle should last quite a while. It absorbs nicely into the skin without feeling greasy or sticky afterward. I also really appreciate the clean ingredients and the fact that there are no added perfumes or strong scents, which is important for my sensitive skin and overall preference for non-irritating skincare products.
As with most skincare products, I know it takes consistency and several months of use before you can truly judge long-term results and improvements in your skin. Because of that, I cannot fully speak yet on how effective it is overall, but so far I have been enjoying using it and have had no irritation or issues. For the affordable price point and quality ingredients, I think this is a great skincare product, and I plan to continue using it and update my review later on with long-term results.
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Reviewed in the United States on May 24, 2026
★★★★★ 4
The Ordinary Hyaluronic Acid 2% + B5 (with Ceramides)
Size: 1 Fl Oz (Pack of 1)
The Ordinary’s Hyaluronic Acid 2% + B5 with added ceramides is a hydration powerhouse that delivers noticeable moisture and placid comfort to dry, dehydrated, or combination skin. Hyaluronic acid draws water into the skin’s surface layers, helping skin feel plump, soft, and smooth, while vitamin B5 supports enhanced surface hydration and recovery.
Adding ceramides is a smart touch because they help reinforce the skin barrier, locking in moisture and preventing water loss throughout the day. The result is a formula that doesn’t just sit on the surface — it helps improve the skin’s resilience over time.
The texture is lightweight and gel-like, absorbing quickly without leaving a sticky or heavy feel, making it easy to layer under serums, moisturizers, and sunscreen. It works beautifully in both morning and evening routines.
After consistent use, many users notice better overall hydration, smoother fine lines caused by dryness, and a more comfortable complexion — especially during dry seasons or in air-conditioned environments.
Pros:
✔ Deep hydration that lasts
✔ Lightweight and fast-absorbing
✔ Great paired with other products (moisturizers / creams)
✔ Ceramides help support the skin barrier
Cons:
✖ Not a direct treatment for acne or pigmentation
✖ Effects build gradually with consistent use
Overall: A very effective and versatile hydrator that’s easy to use and works well for most skin types. Particularly great if you want lasting moisture and a smoother, healthier-feeling complexion.
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Reviewed in the United States on January 27, 2026