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Description
Mouse CYP1A2 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 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 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 P450 1A2 (CYP1A2) 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 P450 1A2 (CYP1A2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Cytochrome P450 1A2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Cytochrome P4501A2 (CYP1A2) is a member of the CYP450 mixed-function oxidase system and a member of the cytochrome P450 superfamily of enzymes. It catalyzes numerous reactions involved in drug metabolism and the synthesis of cholesterol, steroids, and other lipids. CYP1A2 is localized to the endoplasmic reticulum, and its expression is induced by several polycyclic aromatic hydrocarbons (PAHs), some of which are found in cigarette smoke. Other xenobiotic substrates of this enzyme include caffeine, aflatoxin B1, and paracetamol (acetaminophen). Transcripts of this gene contain four Alu sequences in the 3' untranslated region, flanked by direct repeats. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.6 ★★★★★
Based on 22 reviews
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Product Reviews
★★★★★ 5
Good product
Size: 1/2inch x 100ft, Color: White
Good product
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 19, 2026
★★★★★ 5
Works well
This stuff is pretty great I bought it for attaching tomato plants to the fence where the stems were too big to fit my smaller clip ties and even in our strong winds none of the ties have come undone so far
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Reviewed in the United States on June 23, 2025
★★★★★ 5
Effective and functional air purifying
Style: Medium Room Air Purifier, Style: Medium Room Air Purifier
This Amazon Basics model AC2224A Air Purifier is a really capable machine with a somewhat unique filter. I ordered it when it was on special and decided to give some detail on the features to help folks who might be new to air purifiers. The filter weighs in at just over 1lb. That's actually pretty heavy for the size. The filter does the standard 3 things people expect, pre-screen, hepa, active carbon adsorption, but it's a little unique in how it is configured. The pre-screen is straight forward and permanently attached to the outside. It should catch big stuff like pet hair to help stretch out the more expensive layers. Most other filters run a single layer of hepa then separately on the inside run a layer of activated carbon - either as mesh or pellets. Here the filter stitches the hepa and the active carbon together. The active carbon is actually small chunks about the size of sugar crystals layered inside. (yes, I found the end and cut off a pieces to check it out). That actually is a very thoughful way to do it. Activated carbon is structurally changed in the factory to increase its surface area. It's a process you can't replicate at home. The activation allows it to adsorb molecules that cause odor and VOCs. I've given you an illuminated photo of the inside of the filter so that if you can zoom in you'll see the random strands which the hepa standards require. Random strands improve filtering and add strength for the filter. The risk with running a layered filter like this is that if it's constructed poorly it will block airflow. I can check those aspects and on high speed the machine pulls under 30watts and puts out just shy of a 9mph air flow. That's good work. Air flow does create air noise which I measured at 65db directly on top of the machine. It drops another 10db a foot away. On the sleep mode I get 42db on top and that actually drops below the 30db I can measure once you're a foot away from the machine. So, it's capable of power and of being quiet. The last thing is the automation. There is an air sensor on the back by where the cord plugs in that measures the PM2.5 levels. I know what mine are (I have multiple purifiers and sensors), so it doesn't surprise me in the least that on auto the machine drops down to the lowest level. You can also turn on a light that then adds a color to indicate the air purity. I use either essential oil or incense to test the sensor. This was very responsive and noted the essential oil mist I was diffusing right away. It turned the light red and sped to its highest level. Once it had cleaned up the issue it slowed back down and returned to its peaceful green color. It gives solid performance that is in no way basic. Plus, there's the added advantage that you're familiar with the company behind it.
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Reviewed in the United States on January 12, 2026
★★★★★ 5
Fresh Air, Quiet Performance, and Easy Setup
Style: Medium Room Air Purifier
Great air purifier! It was super easy to set up and started working right away. The noise level is low, even on higher settings, and the air in my room feels noticeably fresher. It’s compact, looks nice, and fits well in smaller spaces. Definitely a solid buy if you want something simple and effective.
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Reviewed in the United States on May 1, 2026
★★★★★ 5
Good air purifier - make sure you actually unwrap the filter lol
Style: Medium Room Air Purifier
I'm an idiot and didn't realize the filter that comes already inside it was in there wrapped in plastic for some reason that surpasses my understanding (the whole unit was already wrapped in plastic, why is the filter, which is inside the plastic-wrapped unit, also wrapped in its own separate plastic?? As if we need more plastic in the world...). Anyway, I didn't look inside cuz I thought the filter would just be installed in there and I plugged it in and cheerfully ran it for months til it told me it was time for replace the filter then bought a second filter and went tk take my old one out only to realize I hadn't been filtering anything all that time 😅 oops. Lol. So yeah, make sure you unwrap the filter. Otherwise it's a good little air purifier. Now that the filter is actually installed right, I notice it kicks up higher when we cook or do anything that would lower air quality, so it does seem to be detecting that properly! My allergies do seem better overall. It's pretty quiet when it's on low, but does get loud when it kicks up. I'm OK with that since it pretty much only happens when we cook or if I just put out a candle or something.
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Reviewed in the United States on May 3, 2026