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Description
Human USP6NL 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 ultrasonically disrupted or repeatedly frozen and thawed. Finally, the homogenate is centrifuged at 5000×g for 5-10 minutes and the supernatant is collected 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 conjugate are sequentially added to microwells pre-coated with a capture antibody against USP6 N-Terminal Like Protein (USP6NL). 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 USP6 N-Terminal Like Protein (USP6NL) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human USP6 N-Terminal Like Protein ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | USP6 amino-terminal-like protein (USP6NL), also known as UUSP6 N-terminal-like protein, is a protein encoded by the USP6NL gene. It acts as a GTPase activator for RAB5A and RAB43. It is involved in receptor trafficking. In complex with EPS8, it inhibits EGFR internalization. It participates in retrograde transport from the endocytic pathway to the Golgi apparatus. It participates in the transport of Shiga toxin from early and recycling endosomes to the trans-Golgi network and is required for the structural integrity of the Golgi complex. | |||||||||||||||||||||||||||||||||
| 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 and other biological fluids |
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4.7 ★★★★★
Based on 1861 reviews
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Product Reviews
★★★★★ 1
Cheap, screw holes don't line up, screen is to short, and bars take WD-40 and a mallet
This has been my worst purchase on Amazon ever. DO NOT BUY. My first issue was that bars #4 and #5 would not go together, I ended up buying WD-40 and a mallet and that kind of got them together. Now I went to but the screen on (which is almost see thought) and the screen is to short. I can't attached the bottom bar to the tall bar. The screw hole is in the wrong spot, so the screen is not secure. No amount of pulling on the screen will make it grown .25" longer. Since I can't get the bars secure the walls are not steady and fall over easily. The feet did go together very nice. I can't return them according to Amazons policy. I bought 2 sets. I'm not going to try to build the other set I have. And I can't take apart the first set, since the bars are so tightly together. Also the polls have rust on them.
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Reviewed in the United States on December 10, 2023
★★★★★ 3
Very Poor Built Quality
Color: Black, Size: 3-Panel
They use cheap tarp like cloth that you might find on a tent, so it is not suitable as a teleconferencing background without blur applied. The legs are flimsy and lack rubber feet, so it will scratch floors. The tubing is flimsy and very hard to move and adjust. This is only good if you are setting it up somewhere like a gym or a garage.
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Reviewed in the United States on January 12, 2025
★★★★★ 5
Great separator for room.
My kids love it. It separates their half of the room.
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Reviewed in the United States on February 25, 2025
★★★★★ 5
These are PERFECT for my Art Show coming up!
This is going to be PERFECT for my Art Show display. And also....EASY to put up and take down as well as store. I'm so grateful to have found them! They are super light weight. And sturdy for my application. And elegant and have clean lines for my display. I have ivy draped and also fairy lights which make it look classy yet simple!
Love them!
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Reviewed in the United States on September 27, 2022
★★★★★ 5
Versatile Room Divider With Easy Assembly and Strong Coverage
Size: 3 Panel 12FT W
I picked up this Siebwin 3-panel folding room divider mainly for privacy and room separation, and overall it works very well.
Assembly is very straightforward, and the divider can be set up, taken apart, and stored without much effort. The fabric quality feels good, and the frame construction is stronger and more stable than expected. The support tubes especially feel well built and help keep the divider standing securely.
One feature I really liked is the flexibility of the design. The panels can be used together as a complete divider or separated depending on the setup and available space. That makes it much more versatile for different room layouts or temporary privacy needs.
The coverage is also very good, and the size matches the manufacturer’s description accurately. The fabric blocks light and background visibility well enough to provide solid privacy without feeling overly heavy. The wider feet also help improve stability compared to thinner folding dividers.
Another positive detail is that everything arrived complete with no missing parts or damaged pieces, which made assembly much easier and faster.
Compared to cheaper privacy screens, this one feels more durable and easier to customize depending on the situation. In terms of value for the money, it feels like a very practical and worthwhile purchase considering the size, flexibility, and build quality.
Overall, a versatile and well-built room divider with easy assembly, strong privacy coverage, stable construction, complete included parts, flexible panel configuration, and excellent everyday functionality.
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Reviewed in the United States on May 27, 2026
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