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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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4.8 ★★★★★
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Product Reviews
★★★★★ 5
Nice and comfy, light duty and feel great.
Size: X-Large, Color: Light Green
What they may lack in "durability" i.e. very light and thin = not going to last all that long, they make up for in fit [yes even for wide shoulders], comfort and style. If you want them to last, suggest a gentle cycle and just take ot easy on them. They are light, thin, and airy. Nice summer shirt.
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Reviewed in the United States on May 23, 2026
★★★★★ 4
A true vacation find!
Size: Medium, Color: Light Green
Really nice choice. Got it for my husband and he really liked the style. I loved how it fit on him and the lightness of the fabric. Great for hot, sunny weathered days, or warm nights. A true vacation find. Both comfy and stylish. He would have preferred it be 100% cotton, but I think he didn't really mind after steaming it. He commented on the color too. A beautiful sage green!
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Reviewed in the United States on April 4, 2026
★★★★★ 5
Well made, comfortable and flattering
Size: 3X-Large, Color: Light Green
Nice material. I ordered the 3XL because my hubby has a big belly. It fit him well and was slimming. He wore it as a guest to an outdoor wedding. He looked nice and was comfortable all night.
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Reviewed in the United States on May 6, 2026
★★★★★ 5
Nice but short
Size: X-Large, Color: Lavender
Yes, I purchased this shirt for my husband to wear on Mother’s Day. He looked so handsome and the color complimented his skin tone beautifully. The only con that my husband expressed was that the shirt was short but was comfortable and the fabric felt good on his skin. He is 6’3” and usually wears an XLT. Coofandy, Please make these shirts for taller, longer torso men!
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Reviewed in the United States on May 19, 2026
★★★★★ 5
Expensive Look Without The Expense!
Size: Medium, Color: Black, Size: Medium, Color: Black
Sizing:
If you’re a true large like me and close to but less than fifteen pounds overweight, normal length and with a 44” chest, buy a medium and use the shirt to inspire you to lose the beer belly. If you have similar dimensions but are overweight by up to thirty pounds, get the large.
Materials: The mediums are manufactured to 70% synthetic, 30% linen—which is a great mix, as is doesn’t have to be ironed after it’s washed and hung to dry, nor does it have to be ironed every time you get up from a chair. I’ve had many 100% linen shirts and sure, they look great after you spend thirty minutes ironing them, just to look like a crumpled up newspaper after you get out of your car. My experience with linen shirts is what brought me here.
Unboxing the shirt:
I threw it on an ironing board the moment I unbagged it. I tried the Synthetic setting on the iron—it wasn’t enough. I then tried the Wool setting and set the steam setting to constant medium to high. It worked great. I didn’t have to press down at all, just lightly glided the iron across wrinkled areas of the shirt. Within five minutes, the shirt looked like a hundred-dollar shirt. The material is light, yet feels heavy to the touch and breathes very well in the heat. The hidden buttons give a clean, modern look that allows the shirt to be worn with anything from shorts and sandals to dress slacks and shoes in a club. I like to wear these shirts, untucked with a pair of jeans and western boots. This shirt is very comfortable, fits nice and is extremely versatile.
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Reviewed in the United States on April 5, 2026