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Description
Human COX2 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 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 | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Cytochrome C Oxidase Subunit II ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Cytochrome c oxidase subunit II (COX2), also known as COXII, COII, or MT-CO2, is a protein encoded by the MT-CO2 gene. It is the second subunit of cytochrome c oxidase and one of three mitochondrial DNA (mtDNA)-encoded subunits (MT-CO1, MT-CO2, and MT-CO3) of respiratory complex IV. It is a component of the mitochondrial respiratory chain, catalyzing the reduction of oxygen to water. It is one of three subunits responsible for forming the functional core of cytochrome c oxidase. | |||||||||||||||||||||||||||||||||
| 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 23 reviews
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Product Reviews
★★★★★ 5
DIEGO'S FAVORITE OUTSIDE BALL!
Color: Assorted, Size: Medium
JW PET COMPANY HOL-EE ROLLER EXTREME
This is the second HOL-EE ROLLER ball we have purchased for our Diego. He LOVES these! He now owns a black one and a blue one. He loves to toss these around the yard all by himself or have us throw them to him. He looks so adorable when he comes running back to us, with the HOL-EE ROLLER in his mouth and he has the sweetest expression on his face and is so happy he is prancing! He is a rescue Pit mix from PR. He survived Hurricane Maria and was there before, during, and after. If he could only tell us his tale.
The ball is sturdy. The honey comb design gives your baby the ability to grasp and hold the ball. Diego also likes to hold the ball down and gnaw on it. The ball has proven to be safe against his little vampire teeth. The blue ball was in our yard the entire winter during every type of element you can think of. It is is great shape. Diego plays with these balls constantly while outside. They are sturdy and he is able to get a nice grip on them. These balls are tested by the company to make certain they do not contain any harmful chemicals. The ball has plenty of bounce to it too!
The balls come in different colors but which color you receive will be a surprise.
JW products are AWESOME. Diego also has a trio of smaller indoor balls that he has had for three years now. They are still like brand new. He loves to play with these. They have grips on the outside for him to try to chew on, but he can't destroy them.
JW has other types of pet toys and products available for you. Check them out. Your dog will thank you.
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Reviewed in the United States on June 29, 2020
★★★★★ 4
Most favorite
Color: Red, Size: Medium
My fur babies love these balls they are their favorite. Any size dog can play with it because of how it's made. it easy to play with. It's made with a lot of opening and is made of a soft rubber. My Shepherd pup and Boston's love to play with it, and I had to buy 3 so they don't fight over it. The giggler will break easy and even the first day on rough riders, but they can still enjoy the ball itself even without the giggler. It size is about 4in. If Amazon gave halves Id give it a 3.5 because of the giggler breaking so easily but otherwise my girls still love this ball.
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Reviewed in the United States on May 13, 2026
★★★★★ 5
My boy loves it!
Color: Red, Size: Medium, Color: Red, Size: Medium
This is my frenchies new favorite toy!! He loves the sounds and it seems very durable. Would recommend!
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Reviewed in the United States on May 11, 2026
★★★★★ 5
Wonderful toy, but for most dogs it's a toy you play WITH dog, not a chewie
Color: Red, Size: Medium, Color: Red, Size: Medium
I'm a dog trainer, and have shared my life with many dogs. For the current mini aussie, toys are a necessity and a lifeline. He is tough on toys, and we have certainly tossed many many MANY ruined toys. This toy is wonderful: the squeak is fabulous, it bounces great, it's easy to carry around, and great for fetch and a little tug. Love it, and so does he. But it's soft rubber, so it's a toy to be played JOINTLY with... not a chewie. It will create a lot of fun times and silly entertainment and exercise, but I will put it away when I am not closely monitoring and involved with the play. One of the photos is blurry but shows that the rubber is soft.
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Reviewed in the United States on January 9, 2026
★★★★★ 3
Stops giggling after a few weeks
Color: Red, Size: Medium
I have bought three of these Giggler toys. The dogs absolutely LOVE this giggler toy and it's irresistible to them, even more so than squeaky toys. The rubber is soft and flexible, and after weeks of tug-o-war between dogs, the rubber has stood up well and is showing no sign of breakage. However in two of the toys, the giggler mechanism has started to fail and barely makes any noise at all. The dogs have lost interest in them.
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Reviewed in the United States on August 15, 2024