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Description
Human CYBb 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 Cytochrome b-245 Beta Polypeptide (CYBb) capture antibody. 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 Cytochrome b-245 Beta Polypeptide (CYBb) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Cytochrome b-245 Beta Polypeptide ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | NADPH oxidase 2 (NOX2), also known as cytochrome b (558) subunit beta or cytochrome b-245 heavy chain, is a protein encoded by the NOX2 gene (also known as the CYBB gene). This protein is a superoxide-generating enzyme that forms reactive oxygen species (ROS). It is the catalytic, membrane-bound subunit of NADPH oxidase. It is inactive until it binds to the membrane-anchored p22phox, forming an isoform known as flavocytochrome b558. Upon activation, the regulatory subunits p67phox, p47phox, p40phox, and a GTPase, typically Rac, are recruited into a complex to form NADPH oxidase on the plasma membrane or phagosomal membrane. | |||||||||||||||||||||||||||||||||
| 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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4.8 ★★★★★
Based on 25 reviews
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Product Reviews
★★★★★ 5
Incredible Deal! I bought 2 full suits and absolutely love them!
Size: Medium, Color: Grey
Great looking suit! True to the American Size Chart! I got a lot of compliments the very first day of wearing it! I feel lucky, the Medium Fits like it was tailored! 5’11 (6’ with shoes on, ladies) 190lb. Stocky build. The stretchy waist band is a game changer fellas! Love that! Just like in the video they’re very fun to dance in! The Medium is a lil snug in areas for me but I am currently dieting etc, so if you don’t wanna worry I’m sure the next size up for you fits just as good and will have more breathing room for your arm muscles, thighs, mid section etc as the entire material itself is not stretchy, just the waistband which is incredible!. Overall these are a GREAT score at an amazing price! Perfect for that special occasion or for upgraded business attire! Great Quality! Great Value! and You’ll feel great about yourself in these when you find your faves!
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Reviewed in the United States on February 21, 2026
★★★★★ 1
Packaged in aWrinkled Ball and Bad Sizing
Size: X-Large, Color: Black
I've tried 3 of these suits, 2 different sizes, and each one fits differently. Each one has also been crinkled up and wrinkled each time I opened it. I wouldn't be surprised if it was returned and sent right back out without inspection.
Other reviews noted order a size up. I have a size 34 waist and normally order Large, so I ordered an XL. XL barely can be zippered up. So I returned it. Ordered and XXL. XXL is about 6 inches way too big and took 2 weeks to arrive. So I order and XL again, and once again, way too small, but still bigger than the first..
There is no tag on the pants, only the jacket, so I think someone screwed up the sizing or matching the jacket with the pants.
Total waste of time with 3 returns. Label your products right and get your sizing chart right. There should not be an 8 inch difference between XL and XXL. Got another trip out to UPS, thanks...
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Reviewed in the United States on January 21, 2021
★★★★★ 4
Hubby looks good in it!
Size: Large, Color: Black, Size: Large, Color: Black
This suit is very nice! But it doesn’t have the pockets we were expecting. The left chest pocket is sewn shut along with the outside pockets. It doesn’t have 1 or 2 pockets inside the jacket. Hubby looks good in it!
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Reviewed in the United States on March 5, 2026
★★★★★ 5
Great quality, we could've sized up.
Size: X-Large, Color: Black, Size: X-Large, Color: Black
This suit was absolutely stunning! My son looked amazing in it for homecoming. With him being 6'4", the length was perfect, but it was a but too snug in his thighs, as he is a muscular athlete, and he was a bit worried sitting down in it!😂 Nonetheless, it held up well for my huge Samoan baby, and he got many compliments on it. We also found a bow tie to match it perfectly with the same pattern.
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Reviewed in the United States on September 30, 2024
★★★★★ 3
Jackets made too small and too tight
Size: X-Large, Color: Black
My son is 6'2" & 185 lbs and normally wears a 36 in pants. Site recommends a large, but I ordered an xxl based off the reviews. It was way too big in pants and jacket! I then ordered the large and the xl to be on the safe side because prom was fastly approaching. The large was too small! The xl was okay, but not perfect. Somehow it seems they need a suit size in between the xl and xxl, leading me to think they were just sewn wrong. The pants were extra long (good thing for us), they had to be hemmed. He did need a belt, but not by much. He was able to bend and sit with the pants on. The jacket however still runs small in my opinion. It is too tight in the arms, making it difficult to move. It is also on the short side in sleeves and length. It definitely shows when raising your arms. We chose this suit because we THOUGHT it was the pattern of his date's prom dress; turns out it wasn't. But it matched the vest we bought perfectly.
This suit is made of a thin silky material and it runs small. And the pockets ARE useable. You just need to find the starter hole and remove the thread (if closed). I would recommend buying a different suit unless you order it bigger and have the time and money to alter it.
Oh, and it will take a while to get a refund. Suits ship to you fast, but goes back slow. You gotta wait til the company receive the item, then still wait for the refund.
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Reviewed in the United States on May 6, 2026