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Description
Human SIRT5 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 5 (SIRT5) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted 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 Sirtuin 5 (SIRT5) in the sample. The absorbance (OD value) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Sirtuin 5 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Sirtuin (silent mating type information regulator 2 homolog) 5, also known as SIRT5, is a protein encoded by the SIRT5 gene. This gene encodes a member of the sirtuin family, homologous to the yeast Sir2 protein. Sirtuin family members are characterized by a sirtuin core domain, belong to the third class of the histone deacetylase superfamily, and rely on NAD+ as a cofactor for enzymatic activity. SIRT5 is one of three sirtuins primarily localized to mitochondria. | |||||||||||||||||||||||||||||||||
| 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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4.8 ★★★★★
Based on 25 reviews
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Product Reviews
★★★★★ 5
Cooling King-Size Pillows (used on a Queen pillows to tuck the extra in)
Color: Navy Blue, Size: 2 Pack King（20"X 36"）, Color: Navy Blue, Size: 2 Pack King（20"X 36"）
🛏️ Product Review: Cooling King-Size Pillows (used on a Queen bed)
⭐ The Good
• These are king-size pillows, which I actually love on a queen bed because I can tuck the ends into the fabric.
• The fabric feels slick and cool, but you don’t slip off of it.
• They feel cold to the touch right out of the package (which is wild).
• I sleep extremely hot and even run fevers at night due to a medical condition (literally 101–102° sometimes), and these pillows still stay cool where I lay my head.
• I’ve honestly never slept this well in my life.
• For the price, you cannot beat them, and they go on sale often.
• These beat any high-end version I’ve tried.
• If you sleep normal-temp and not lava-hot like me, you’re going to be VERY comfortable with these.
⚠️ The Bad
• The fabric does show crease lines.
• If wrinkles bother you, this might annoy you (doesn’t bother me personally).
🔧 What I’d Change / Tips for Care
• Don’t use fabric softener — it clogs the cooling fabric and ruins how it works.
• Wash with unscented detergent + a little vinegar.
• Dry on low heat only (do NOT dry on high if you want the fabric to keep working long-term).
✅ Final Thoughts
I will literally never not own these again.
If you sleep hot, night-sweat, or just want a pillow that actually stays cool — these are it.
10/10 recommend.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 17, 2026
★★★★★ 3
Love it but it stains
Color: Haze Blue, Size: 2 Pack Standard（20"X 26"）
I love these but had to knock some stars down for a single reason. They are soft, the material is somehow both substantial and airy at the same time, and a smooth, silky (not satin), cozy texture. The color was exactly as pictured. The envelope style works great to keep my pillows covered. The fabric lays perfectly. I love sleeping on this pillow case.
Took two stars off because every single atom of oil on my face stained it immediately and won't come out in the wash. Its not subtle. You can see anywhere my face has been, forever. I don't have some incredibly oily face or hair, I've never had this happen to another pillow case in my life.
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Reviewed in the United States on April 20, 2026
★★★★★ 5
Good buy
Color: Navy Blue, Size: Body（20"X 54"）
Nice and soft , well packaged and looks good. Kinda pillowcase you find in a luxury hotel
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Reviewed in the United States on May 1, 2026
★★★★★ 5
Silky and Soothing
Color: Cream, Size: 2 Pack Queen（20"X 30"）
I thought I was getting 1 of the queen sized but I got 2. I got these for my Pomeranian's fuzzy pillow. He's always hot and these are very cooling and very good quality. He loves them and are very comfortable for him. They totally transformed his shaggy old pillow. I used the larger body pillow covers to replace the covers I had. They matched my bedspread perfectly. I use 2 body pillows along the exterior wall in my bedroom where there is cold coming through the floor in the winter. I'm sure I will enjoy them in the summer when it's hot too!
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Reviewed in the United States on January 10, 2026
★★★★★ 5
So soft and amazing quality
Color: 12 - Light Beige, Size: Twin
I can't tell you if they're cooling or not yet. The weather took a turn for the cold so I haven't put them on the bed yet. But the quality..omg it's exceptional! These are insanely soft! Not slippery like satin but not scratchy like percale. They're like the best of both worlds..light as satin but nicely weighty like percale. I don't know what kind of sorcery this is but these are literally better than those 6-zillion count Egyptian cotton sheets that cost more than the GDP of a small nation. These are exceptional!! Soft as butter but still feel like they'll last a while and not let you fall out of bed like silk lol.
When you fold them, they snap nicely.
They seem to have a lived in look..like they'll never be utterly wrinkle-free but who cares..they're smooth, soft and supposed to be cooling.
I never thought I'd find anything more comfortable than cotton sheets but these are even better.
The colors are ok. Pretty basic but they are true to life..some others look different online than when you get them..not these colors online are accurate.
This was a great deal and I'd recommend a thousand times over!
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Reviewed in the United States on May 23, 2026
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