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Description
Human SIRT4 ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. 2. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. 3. 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 the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL 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 the supernatant can be assayed. 4. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend 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 freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. 5. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 6. 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 a deacetylase sirtuin-4 (SIRT4) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of deacetylase sirtuin-4 (SIRT4) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Deacetylase Sirtuin-4 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Sirtuin-4 (SIRT4) is a mitochondrial protein encoded by the SIRT4 gene. It is a mitochondrial ADP-ribosyltransferase that inhibits the activity of mitochondrial glutamate dehydrogenase 1, thereby downregulating insulin secretion in response to amino acids. Its deacetylation of malonyl-CoA decarboxylase inhibits the enzyme's activity, suppressing fatty acid oxidation in muscle and hepatocytes. It also inhibits peroxisome proliferator-activated receptor α (PPAR-α), downregulating fatty acid oxidation in hepatocytes. Its overexpression inhibits cancer cell proliferation by suppressing glutamine metabolism. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.8 ★★★★★
Based on 11 reviews
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Product Reviews
★★★★★ 4
Works well, long use on single charge, high torque - be careful when starting
Color: Black, Size: Rechargeable
Not the most inexpensive powered whisker bur I've used other Typhor products and found them reliable for the task designed. I fell they don't cheap out on the rechargeable battery so I don't fell they will catch fire, as we are hearing about from other cheaper rechargeable items. I liked 2 things about this model the speed control is by a pressure single switch which I can control, not just pick speed 1,2,3. The swappable tips allow for different uses. I don't drink coffee but use the item to whisk eggs for an omelets and it has the torque to mix them without effort. In Fact need to only use half power else it will splatter out of my mixing vessel.
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Reviewed in the United States on June 7, 2026
★★★★★ 5
I love it!
Color: Black, Size: Rechargeable
This thing works great! It holds a charge for a long time. I love that it has different attachments. I use it mainly for mixing powder into my drinks. It is very powerful. The speed is adjustable too. It feels good to hold too. It lights up when you squeeze the trigger and when it starts losing charge, the light bar gets shorter. I highly recommend it.
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Reviewed in the United States on June 3, 2026
★★★★★ 5
Perfect all-around frother, stand keeps it nice and neat on the counter within easy reach.
Color: Black, Size: Rechargeable
Rechargeable, lightweight, variable speeds and optional whisks all make this the perfect item for your kitchen. You can quickly froth milk for your coffees, whip a small batch of whip cream, and even make mug cakes with the hook attachment. The price is very compatible with the competitors, but the ease of use makes this a top choice.
The variable speeds makes it much better to control than the push button ones. When you let go of the pressure switch it turns off, no need for a dedicated on-off switch. Having the pressure switch on the top makes it extremely easy to use! It fits very comfortably in the hand and with the metal stand, looks quite nice on the counter, ready to use at any time. I highly recommend this InstaWhisk!
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Reviewed in the United States on May 28, 2026
★★★★★ 5
Great product from a great company
Color: Black, Size: Rechargeable
This product works very well. There are several other whips to use also. I absolutely love the cream it makes and that is important so I can enjoy my morning coffee. I used this whisk for quite a while then all of a sudden it stopped working. I wrote to the company to explain. A few days later they sent me a beans new whisk. GREAT item and great company to work with.
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Reviewed in the United States on May 23, 2026
★★★★★ 4
Great Frother - almost too powerful
Color: Black, Size: Rechargeable
I’ve been using the Tyohur InstaWhisk milk frother for a while now, and overall I really like it—but I wouldn’t quite give it 5 stars, and honestly it’s because it might be a little too powerful for its own good.
First, the positives: this frother does an amazing job. The single-button control is super easy to use, and I love that you can adjust the speed just by how much pressure you apply. It feels very intuitive, and you can really dial in the exact texture you want. Whether I’m making a latte or mixing something thicker like a protein drink, it handles everything smoothly.
The performance is seriously impressive. On the lower speeds, it creates nice, silky foam, and when you turn it up, it’s incredibly fast—like café-quality foam in seconds. High-speed frothers like this are designed to create rich foam very quickly, and this one definitely delivers on that promise
That said, the power is also what knocks it down a star for me. If you’re not careful (especially when using higher speeds), it can splash pretty easily. The product description says it’s designed to prevent mess with stable low-RPM operation, and that’s true if you stay on the lower settings—but once you ramp it up, you really need to keep the whisk fully submerged or start slow, or it can send milk flying out of the container
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Reviewed in the United States on May 23, 2026
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