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Description
Rat COX2 ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. 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 the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL 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 the supernatant can be assayed. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend 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 freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 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 utilizes 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 C Oxidase Subunit II (COX2) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of Cytochrome C Oxidase Subunit II (COX2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Cytochrome C Oxidase Subunit II ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Prostaglandin endoperoxide synthase 2, also known as cyclooxygenase 2 or COX-2, is an enzyme encoded by the PTGS2 gene. It is one of two cyclooxygenases involved in the conversion of arachidonic acid to prostaglandin H2, an important precursor of prostacyclin expressed in inflammation. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.3 ★★★★★
Based on 446 reviews
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Product Reviews
★★★★★ 1
FAKE! Caused burning stinging and pain. AVOID
Counterfeit product-avoid! Don’t buy it from here, get the actual product from Bioderma directly. I used this and it made my face sting, itch and turn a bright red. Then came the BURNING! After avoiding it for a week my face is finally healing. I didn’t notice the horrible packaging until it was too late—compare the photo of the product I got to the actual product from Bioderma and you’ll see the difference.
It is extremely unethical for a seller to send FAKE products that cause PAIN and redness all to make a dollar. Not cool.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 15, 2026
★★★★★ 5
Good
Good
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Reviewed in the United States on May 25, 2026
★★★★★ 4
Cumple con las expectativas
Crema ligera de rápida abosrcion. No sé percibe muy rápido que calme el enrojecimiento de la piel
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 20, 2026
★★★★★ 5
All good thanks
All good thanks.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 17, 2026
★★★★★ 5
I Just Want to Help People With Skin Problems!
Style: Tolerance Control Soothing Skin Recovery Cream
I so wish I had known about this skincare line a long time ago, but just glad I did find it at all. I have rosacea (including ocular rosacea) diagnosed by a dermatologist. I had been through all the typical stores and websites, trying so many things over the last 15 years. Yes, most were supposed to be for people with skin problems. EVERY SINGLE TIME I tried something, within five minutes, my skin was bright red. My skin reacted to EVERYTHING. I started to believe there was not one single product on the entire planet that my skin would not react to! It was so frustrating. Purchase, fail, repeat. Thing is, I had tried their spring water spray. I sort of ignored all the other products until I was talking to someone randomly and they asked if I had tried Avene. Honestly I decided maybe this was providence and that I had better try it just to be on the safe side. What would life be like if I had ignored their advice and not tried it? If this company ever goes out of business, I will have to live with terrible skin because I already know there is nothing else out there. I had read a review whereby someone said their doctor told them the longer they used the products, the better their skin would get. This is completely true. But even initially, all I cared was that I could relieve the dryness with SOMETHING that did not turn my skin RED IMMEDIATELY! Even THAT, I would have been happy with! Now for the rest of the benefits. I had the good intuition to purchase two of these when they were on sale. I am so glad I did. I use it like crazy every chance I get. It just feels so good going on and I like touching my skin even after the product dissipates. No redness, no irritation, it actually CALMS my skin. And makes it like, well, "real skin." I'm glad I also got the thicker version in the tube, Cicalfate. This is my go-to for dryer areas and in the winter. The two together along with the water spray, if you like to use something like that, are a great overall program. Definitely other things I would like to try. Again, my goal was to help others who have skin problems because everything else out there has no concept whatsoever of calming the skin, but the exact opposite -- irritating it worse (even if those products are supposed to help with redness or irritation). And I felt the company truly deserves a shout-out because they GET IT when it comes to skincare products. Whatever they've done, they are experts, far more than all the other skincare lines and I don't care how long those have been around. If it doesn't immediately tell you your skin "likes it" and shows it by not reacting or immediately getting better, why on earth would you pay your hard-earned money to buy it and keep using it? It's worth every penny, don't bat an eyelash, your skin is worth that and more. If it's on sale, get a lot of it because trust me, you are going to want and need it. You will be so glad you got more than one. I definitely am! THANK YOU, AVENE, WE LOVE YOU HERE IN AMERICA! YOU UNDERSTAND SKINCARE LIKE NO OTHER COMPANY!
Update: Wanted to comment that I'm still so grateful for a product that does FAR MORE than what you expect! I recently bought more. I watch my skin calm down the minute I put it on. I don't know entirely how the ingredients do what they do, but to the company ... please don't fix what isn't broken! Too many companies have a good thing and then change it. Please don't, the products are perfect and far beyond expectation! I also wanted to mention it seems the Tolerance tube will get EVERY LAST BIT out, you won't be wondering if there is anything left you are discarding. The Cicalfate tube is a bit different, it's more like a squeeze tube. But I feel I can still get the product out to use it all. That said, you get your money's worth. I'm also starting to use the Cicalfate on my hands in the winter, too (which are sadly starting to show my age). But this product line has literally CHANGED MY LIFE! BUY IT, it's the REAL DEAL!
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Reviewed in the United States on August 10, 2025
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