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Description
Human USP11 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 competitive enzyme-linked immunosorbent assay (ELISA). Samples, standards, biotin-labeled antibodies, and HRP enzyme conjugates are added sequentially to microwells pre-coated with the universal species Ubiquitin Specific Peptidase 11 (USP11) antigen. After incubation and washing, the sample is developed with the substrate TMB. TMB is converted to blue under the catalysis of peroxidase (HRP) and to the final yellow under the action of acid. The depth of the color is negatively correlated with the universal species Ubiquitin Specific Peptidase 11 (USP11) in the sample. The absorbance (OD value) is measured at a wavelength of 450nm using an enzyme reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Ubiquitin Specific Peptidase 11 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Ubiquitin-specific peptidase 11 (USP11), also known as UHX1, is an enzyme encoded by the USP11 gene. It belongs to the ubiquitin-specific protease family (USPs), a subfamily of deubiquitinating enzymes (DUBs). USPs are multidomain proteases belonging to the C19 cysteine protease subfamily. Different members exhibit varying homology based on their domain architecture and location. It is a 963-aa protein with a molecular weight of approximately 109.8 kDa and a pI of approximately 5.28. It shares significant homology with USP15 and, together with USP4, forms the DU subfamily. Protein ubiquitination controls many cellular processes, including cell cycle progression, transcriptional activation, and signal transduction. This dynamic process involves ubiquitin-conjugating enzymes and deubiquitinating enzymes, which add and remove ubiquitin. Deubiquitinating enzymes are cysteine proteases that specifically cleave ubiquitin from ubiquitin-conjugated protein substrates. This enzyme is located in a gene cluster on chromosome Xp11.23. It has been shown to interact with RANBP9. | |||||||||||||||||||||||||||||||||
| 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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4.5 ★★★★★
Based on 790 reviews
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Product Reviews
★★★★★ 5
Best air flow
Size: 9.6" x 11.1" x 2.2"
I have a 2022 ram 1500 four-door. The factory filter from the ram dealer is about $40 but it’s not reusable. That was the only one I found that had good airflow into the cabin. I tried several other brands but the airflow was reduced excessively. The ram filter was 15 mph, most of the other aftermarkets dropped the airflow to about 9 mph. I just installed a K&N reusable filter and the airflow is back up to 15 mph. Great job K&N!!
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Reviewed in the United States on March 27, 2026
★★★★★ 5
Fits Well and Reusable. I'm Happy.
Size: 9.6" x 11.1" x 2.2", Size: 9.6" x 11.1" x 2.2"
For less than the price of 2 disposable FRAM CV11671 cabin filters for my 2019 RAM 1500 and this will last forever. If it gets wet, like I've had happen with the paper throw-away filters due to stuff falling off trees in the spring and clogging cowl area water drains and backing up into filter ( yeah, that's a thing ) and can clean and dry this filter and reuse. Great directions came in the box along with a sticker to point out that this is a lifetime filter so someone doesn't throw it away. Do this yourself. Don't let some auto repair shop talk you into it. I was getting tires balanced recently and heard a quote they gave over the phone of $79 to do this. I don't know what kind of vehicle that was for but this ain't rocket science. Don't get ripped off. Now back to this filter. There is no arrow for air flow so I figure it flows well both ways and just put it in with the K and N logo right side up and facing out. It's not a straight in shot the way RAM positioned the opening for the cabin filter slot but was no big deal. Also realize this isn't a perfect square filter ...It's a rectangle so don't try to shove the fat end in and say it doesn't fit. I haven't noticed any air flow differences with the heating / ac system but some people say they do. I'm just happy I don't have to buy anymore disposable filters and can dry this if it gets wet. The cleaning and refresh spray kit is #99-6000 that comes with a 8oz and 12oz bottle and I'm trying to find that cheaper somewhere for when I do clean this thing.
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Reviewed in the United States on November 20, 2025
★★★★★ 5
Can’t go wrong with K&N
Size: 9.6" x 11.1" x 2.2"
So I recently took my pickup in to have some recall work done, half way through the process I got a call from the dealership stating I needed a new air filter and that a new one would cost nearly $60. I chuckled and told then to put the old one back in and I immediately came to Amazon and ordered this cabin filter. I’ve used K&N products in the past and have never been sorry, and that is the case with this lifetime filter. Upon installing this filter I noticed an IMMEDIATE difference in air flow into the cabin, it even smelled better. If this filter ever gets dirty Ira easy to clean and reinstall, paying for itself over time. Do yourself a favor and pick one of these up, you will not be sorry.
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Reviewed in the United States on October 20, 2025
★★★★★ 5
Perfect fit
Size: 9.6" x 11.1" x 2.2"
Perfect fit on my 2022 jeep wagoner
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Reviewed in the United States on May 20, 2026
★★★★★ 5
Easy replacement and noticeably fresher air
Size: 1-Pack
Installation was quick and simple, and it fit perfectly in my vehicle.
I noticed an immediate improvement in the air quality inside the car. The activated carbon helps reduce odors and keeps the air feeling cleaner and fresher, especially during everyday driving.
The filter feels well-made and durable, and it does a great job trapping dust and debris. It’s a simple upgrade that makes the driving experience much more comfortable.
Overall, this is a great quality cabin air filter that delivers reliable performance and excellent value. Definitely recommend!
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Reviewed in the United States on May 8, 2026