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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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4.2 ★★★★★
Based on 17 reviews
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Product Reviews
★★★★★ 5
Strong coffee smell. Might not be be compatible for everyone. However, product is really good.
Scent: Smoky coffee, Size: 4 Fl Oz (Pack of 1)
Love this product. May not be compatible for someone who doesn't like smell of coffee that does not detract from the product. it's really good for hair.
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Reviewed in the United States on January 5, 2026
★★★★★ 3
So far nothing amazing
Scent: Smoky coffee, Size: 4 Fl Oz (Pack of 1)
So I’ve been using for a couple of weeks now and I haven’t noticed any difference. I am going to continue on. I don’t know if it takes time for it to actually show some results. I will say two things about it. It comes solid for 12 my jar cracked and broken And it’s non-returnable so I just have to deal with it so that’s unfortunate after paying $20 second of all it is a solid mass so you have to put it in your hands and kind of warm it up for it to become any sort of an oil and I wouldn’t even consider it an oil at that it’s more of like a cream once you warm it up in your hands, but it doesn’t make your hair greasy. I use a small amount I would think that the jar would last a long time given that you don’t need a lot to get through to your scalp unfortunately, with my jar being cracked, I don’t know how that’s gonna affect the longevity of the product And it does have a smell think of burnt coffee beans that will dissipate after about an hour to an hour and a half after putting it on, but if you’re not putting it on, I’m just going to bed. It might be become an issue. I’m gonna wait it out. I hope I do see results from doing this but again so far nothing.
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Reviewed in the United States on February 15, 2025
★★★★★ 5
Wonderful product
Scent: Smoky coffee, Size: 4 Fl Oz (Pack of 1)
I used this on a bald spot I had and it worked extremely well. Consistency is key though it needs to be used every day
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Reviewed in the United States on April 3, 2026
★★★★★ 5
It works and I love it!
Scent: Smoky coffee, Size: 4 Fl Oz (Pack of 1)
I love this product.. been using it for a few months now. My hair is more manageable and my thinning crown looks and feels fuller. The smoky smell, am not bothered because it disappears after a few minutes. I use it twice a week, massage very little amount on scalp and hair, then the next day wash it off with shampoo. I used to need to dye my gray every 3 to 4 months when some appear. Now, it's been 4 months since my last visit to the salon. I noticed my hair has darken a bit. I can delay my salon visit for another month unless I need a haircut!! Yehey!
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Reviewed in the United States on September 19, 2025
★★★★★ 3
Good moisturizer. Rich scent.
Scent: Smoky coffee, Size: 4 Fl Oz (Pack of 1)
Raw Batana Oil provides goods moisturizing to hair and scalp and best applied with a little heat or steam. Pomade goes on easily once heated and does not feel heavy, greasy, or oily. Its rich fragrance eases on the nose after a short while. Difficult to say whether it is actually useful or whether it encourages hair growth. Leaves hair soft and easy to style.
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Reviewed in the United States on December 28, 2025