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Description
Human PIAS3 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 utilizes 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 capture antibody against the E3 SUMO-protein ligase PIAS3 (PIAS3). 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 E3 SUMO-protein ligase PIAS3 (PIAS3) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human E3 SUMO-protein ligase PIAS3 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Protein inhibitor of activated STATs 3 (PIAS3), also known as the E3 SUMO-protein ligase PIAS3, is an enzyme encoded by the PIAS3 gene. PIAS3 was initially discovered to interact specifically with phosphorylated STAT3 in interleukin-6 (IL-6)-activated myeloblast M1 cells. This interaction is mediated by PIAS3 binding to the STAT3 DNA-binding domain. Consequently, STAT3 transcriptional activity is inhibited by physically preventing its binding to target genes. Subsequently, it was also discovered to be a regulatory protein of other key transcription factors, including MITF, NFκB, SMADs, and the estrogen receptor. It also functions as a SUMO (small ubiquitin-like modifier)-E3 ligase, catalyzing the covalent attachment of SUMO proteins to specific target substrates. It directly binds to several transcription factors and either blocks or enhances their activity. Alternatively spliced transcript variants of this gene have been identified, but the full-length nature of some of these variants has not yet been determined. | |||||||||||||||||||||||||||||||||
| 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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4.7 ★★★★★
Based on 17 reviews
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Product Reviews
★★★★★ 5
Serves its purpose
If you can get past the texture of the silicone these will work wonders for your feet. There’s no odor, easy to put on but they are cut small. There’s no moisture it just feels moist because of the silicone. They are very soft and durable.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 11, 2026
★★★★★ 3
Dissatisfied
Didn't like the socks at all. The silicone in them made my feet sweat and couldn't wiggle my toes at all.
Gloves were okay, received them with holes between the fingers, only able to wear each twice.
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Reviewed in the United States on November 17, 2025
★★★★★ 2
too small
I saw these and instantly thought they would be amazing! I was hoping for just that. I received them today and tried them right away. I love the gel feel but im not really liking the way they kept rolling off. They're a bit too small id say and when walking they roll off from the back. I didnt think my foot was other than average? Maybe? Im a size 7 in women's shoes.
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Reviewed in the United States on September 12, 2025
★★★★★ 5
Super Awesome
I have the driest feet, my mom did too so I should have known that I would inherit it as an adult myself. My toes literally crack and split open from dry skin, I call them fissures. They really hurt. Last night I used my cheese grater foot file gently on my heels and calloused parts, and then applied liberal lotion, and wore those squishy socks all night. I woke up to the most moisturized youthful feet I’ve ever seen attached to my body! Wow these totally work!!! I even slept better than I have in a very long time. I’m very pleased. These did exactly what I hoped they would do.
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Reviewed in the United States on February 18, 2026
★★★★★ 5
Amazing!
Number of Items: 1
Wow, what to say about this glitter spray. If you want to cosplay as a Twilight vampire, buy this spray. Haha. I’m not even kidding. I got this spray for going to a concert - one of the few occasions I go all out in getting dressed up. I LOVE sparkly/shiny/glittery things, so this spray seemed like an obvious choice. I did a very light, quick spray over my face/neck and OH. MY. GOSH. “I look like a vampire from Twilight.” My first thought/words after looking at myself in the mirror. My mom got a pretty big kick out of it and couldn’t stop giggling because that’s all she could think about afterward whenever she looked at me.
I expected this spray to be something I’d have to spray multiple coats of, but absolutely not. This is a one and done spray. The glitter is HIGHLY CONCENTRATED. Very, very sparkly with multifaceted color sparkles - not just silver. I got some on my hair when I sprayed myself, so I went ahead and sprayed the back of my hair too. Didn’t put any kind of film or anything in my hair. I was still able to brush it and do everything else as if there was nothing in my hair. At the end of the night, it washed off very easily.
I will say as a pointer, unless you want your hair or clothes to have glitter on them as well, then you should apply this glitter spray BEFORE you get dressed and you should wear a shower cap/bag/something to cover your hair while applying. However, it works just fine on your hair and clothes as well - no staining or damage.
The can looks small BUT with how little you have to use, one can will go a very long way. I could go to 10 concerts and I’m pretty sure this can would still be more than half full.
HIGHLY RECOMMEND!
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Reviewed in the United States on April 13, 2025
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