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Description
Human 5-LOX 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 conjugate are sequentially added to microwells pre-coated with a 5-Lipoxygenase (5-LOX) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of 5-Lipoxygenase (5-LOX) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human 5-Lipoxygenase ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Arachidonic acid 5-lipoxygenase (ALOX5, 5-lipoxygenase, 5-LOX, or 5-LO) is a non-heme, iron-containing enzyme encoded by the ALOX5 gene. It converts essential fatty acid (EFA) substrates into leukotrienes and a wide range of other bioactive products. The ALOX5 gene occupies 71.9 kilobase pairs (kb) on chromosome 10 and consists of 14 exons and 13 introns, encoding the mature ALOX5 protein of 78 kilodaltons (kD) composed of 673 amino acids. | |||||||||||||||||||||||||||||||||
| 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 | 0.15-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.0 ★★★★★
Based on 27 reviews
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Product Reviews
★★★★★ 5
Very comfortable Slippers. Great buy!
Size: 12-13 Women/10.5-11.5 Men, Color: Charcoal
I bought these to replace a pair of slippers that I have had for years that are very similar to the slippers I bought, but I had worn a hole in the leather on mine, so I had to find something new and similar. These slippers are worth the money they are very comfortable, and I am sure they will last for years. They are a little tight around the ankle cuff but they are still new and I am sure they will wear in a little more when I start wearing them more.
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Reviewed in the United States on March 8, 2025
★★★★★ 3
Great slippers but disappointing quality.
Size: 7.5-8.5 Women/6-7 Men, Color: Ash, Size: 7.5-8.5 Women/6-7 Men, Color: Ash
Extremely comfortable, attractive slippers but the stitching came loose on both leather trim areas almost immediately. I tried to sew/glue down the stitching (as seen in photo) but it kept unraveling. Disappointing as these slippers are great in every other way (though they do run a bit big...I wear a 9 but ended up with a 7.5-8.5 which fits with a bit of room to spare even with thick socks). I just hope they hold up after my latest attempt to stop the thread from pulling out any more as I do really love wearing them.
Perhaps I just got a bum pair?
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Reviewed in the United States on April 3, 2024
★★★★★ 5
Tight at first, but they stretch after a short time...feel perfect after the first week!
Size: 9.5-10.5, Color: Charcoal Faux Fur
I love them!
My first impression was "oooh, this is too tight". I wear a size 10 shoe and honestly, the 9.5-10.5 felt tight when I first tried it. Without socks it felt okay, with socks my toes were right at the top. But I loved the style so much I decided to give it a chance - I am SO glad I did!!!
They do stretch and give, so don't expect it to be tight forever. I have had them a week or so, wearing each day (I WFH) and they feel really nice. I also live in the mountains, we just had our first winter storm, and these slippers are nice and toasty. Exactly what I needed.
The memory foam is plush, though I have had other slippers with it and I know it will squash down over time. Right now, it is a nice feeling as I walk around the house. They are super cute looking, wearing leggings allows me to feel like I am stylish even while still wearing my sleep thermals. YAY!
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Reviewed in the United States on November 18, 2025
★★★★★ 5
favorite slipper ever
Size: 8-9, Color: Charcoal Faux Fur
I love these slippers. They look great with pants tucked in. Incredibly warm and comfortable. I wear them almost every day. Fit is true to size. They stay up but are not tight at all. On top of that I get compliments on them. Highly recommend.
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Reviewed in the United States on May 3, 2026
★★★★★ 5
Favorite Slippers Of All Time
Size: 6.5-7.5, Color: Charcoal Faux Fur
ETA 7/2024: So I ended up buying a size 5-6. They fit better around the top and sides of my foot but the sole is a bit small. The 6.5-7.5 sole fits better but the material around the top and sides is roomy so it bunches up. Sigh. I still love these slippers but the fit is frustrating.
I'm obsessed with these slipper boots! They are so cute and comfy.
They look especially good with leggings but I also like that I can tuck other types of pajama pants into them. I'm short and have to fold the waist of most pajama pants so they're not super long on me, but with these boot slippers on I don't have to worry about my pant bottoms dragging on the floor since I can just tuck them in.
I like that they have rubber soles. I love the look of the faux fur but not sure how easy they will be to wash because of it.
I wear a 7 or 7.5 and I got the 6.5-7.5 size. Even with thick socks on they are roomy. Because they are house shoes it doesn't matter much but I probably could've sized down.
I like the way they look so much I might buy a size down and use that pair for wearing outside of my house.
Some other reviews said they have a strong smell but mine did not.
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Reviewed in the United States on January 1, 2024