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Description
Mouse PPARa 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: 1. 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: 1. 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. 2. 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. 3. 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. 4. 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. 5. 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. 6. 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 capture antibody against Peroxisome Proliferator Activated Receptor Alpha (PPARa). After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of peroxidase (HRP) and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of Peroxisome Proliferator Activated Receptor Alpha (PPARa) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Peroxisome Proliferator Activated Receptor Alpha ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Peroxisome proliferator-activated receptor α (PPAR-α), also known as NR1C1 (nuclear receptor subfamily 1, group C, member 1), is a nuclear receptor protein encoded by the PPARA gene. Along with peroxisome proliferator-activated receptor δ and peroxisome proliferator-activated receptor γ, it belongs to the peroxisome proliferator-activated receptor subfamily. It was the first member of the PPAR family cloned by Stephen Green in 1990 and was identified as a nuclear receptor for a distinct class of rodent hepatocarcinogens that promotes peroxisome proliferation. It is primarily activated by ligand binding. | |||||||||||||||||||||||||||||||||
| 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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4.9 ★★★★★
Based on 13 reviews
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Product Reviews
★★★★★ 1
Quit working in a month
Bought one. It quit working. They sent another… it never worked right before finally dying on me the same way. Looks like it’s not even an available product anymore. But if it ever does come back.. don’t buy it.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 25, 2025
★★★★★ 5
Not what I expected!
I have never used a dental flosser before, and it showed by the mess I made haha.
I can say, this isn't for me, but I can see the appeal. It is relatively lightweight and comes with a lot of different attachments. Loading it up with water is easy and doesn't take much effort.
I would call this portable, even though it is a little big.
I can only stand the "soft mode". If I choose any other, my mouth is overwhelmed.
Overall, I would definitely recommend this if you have experience with dental flossers. It was hard to use at first but you get the hang of it.
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Reviewed in the United States on July 1, 2024
★★★★★ 1
Reasonable cost dental device (updated review)
I used this device for a few months to provide an accurate review.
First off, it works. Fill the bottom end section with water, snap in a tip, switch on and go. Easy to use, and the instruction set is short and simple, the result of a thoughtful adoption process. Very reasonable cost. Several heads are provided for you to select. And there are three settings to use: soft, normal and pulse. I did not find any benefit to the pulse setting as it seemed to cycle the water pressure lower than to normal repeatedly.
Several additional items worth mentioning:
1) The water pressure is modest. the lower pressure is made up for in more water delivery.
2) This device consumes water at a rapid pace. What this means is that you will need to either swallow or spill the water out of the side of your mouth at least twice during the 90 second cycle - while the unit is operating. Switch off during the cycle restarts the countdown. Water will accumulate rapidly in your mouth. You need to spill the accumulated water in your mouth with the unit on. If you remove it from your mouth you will have a mess around you.
3) The on-off switch is located on the handle toward the top. You will need to learn to not place your finger over the switch which abruptly shuts the device off. Conversely, if you like to swith off the device during the cycle to neatly spill accumulated water out of your mouth, you may find yourself attempting to find the switch push button without a tactile means of feeling for it.
4) The charge port has a small flexible grommet covering it, which might take a little while to find where it is located.
5) **This is my update from my original posting**: the duration of charge shortened over the last several weeks to the point where with the unit switched on, the lights are on, but the pump no longer works, meaning it died and has been trashed.
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Reviewed in the United States on May 16, 2024
★★★★★ 4
Decent water flosser.
This water flosser is a handy way to clean your teeth, especially when you're on the go. It's cordless and rechargeable, and it comes with different tips for various needs. The water tank holds enough for a full cleaning, and it doesn't leak. It has multiple settings, from gentle to strong, to get your teeth feeling fresh. While the tips might not be the most durable, and attaching the top can be a bit tricky, it's a great tool for keeping your mouth clean and healthy. It's a good alternative to traditional flossing, helping to remove bits and keep your gums healthy.
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Reviewed in the United States on March 15, 2025
★★★★★ 5
They work great!
My son loves his vibrating toothbrush and it needed a new head. We change the head out every time he is sick and he was just getting over Covid. The brush head is high quality, it doesn’t shed and fits perfectly. The cost per brush is excellent! We will definitely be purchasing again.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on August 14, 2024
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