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Description
Mouse CPT1A 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 capture antibody against carnitine palmitoyltransferase 1A, Liver (CPT1A). After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of carnitine palmitoyltransferase 1A, Liver (CPT1A) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Carnitine Palmitoyltransferase 1A, Liver | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Carnitine palmitoyltransferase 1A, also known as CPT1, is encoded by the CPT1A gene. Mitochondrial oxidation of long-chain fatty acids is initiated by the sequential action of carnitine palmitoyltransferase I (located in the outer membrane and detergent-labile) and carnitine palmitoyltransferase II (located in the inner membrane and detergent-stable), along with carnitine-acylcarnitine translocase. It is a key enzyme for carnitine-dependent transport across the inner mitochondrial membrane, and its deficiency results in reduced fatty acid β-oxidation rates. Alternative splice transcript variants encoding different isoforms of this gene have been identified. | |||||||||||||||||||||||||||||||||
| 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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4.5 ★★★★★
Based on 16 reviews
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Product Reviews
★★★★★ 4
Cute and so much fun!
Size: Small
Great squeak noise!!
Our dog loves to play fetch with them!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 7, 2026
★★★★★ 5
An instant favorite for my one year old pup!
Size: Small, Size: Small
I came across these randomly while looking for presents for my one year old 10lb chi-mix. I gave her one to start with just to make sure it wasn’t flimsy & she wasn’t going to destroy it because she is definitely a chewer. She was instantly OBSESSED! These are now her most treasured squeakies. They’re the perfect size for her too and I’m happy to say they’ve held up well to her pouncing/jumping on them constantly. They’re definitely not quiet but we get an absolute kick out of watching the way she plays with them. If I could give 10 stars I would. Almost two months later and the novelty hasn’t worn off to her. I’ve saved them to my lists just in case we ever need to replace them for any reason. I’m not sure she’ll ever love a toy as much as these
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Reviewed in the United States on January 9, 2026
★★★★★ 5
Squeak is not super loud! Family fun!
Size: Small
My small dog loves these toys. He detects them the very moment they enter the house, even more if the package has been opened. I used to give him one at a time because he would favor the newest one for quite a long time. Now he sniffs them out no matter where I store them.
The squeakers have fallen INTO the toy rather than going down his throat, and they're rather difficult to chew out of place. They do collect quite a bit of debris and hair from the floor, so a good rinsing is needed sometimes. My little 13 lb terrier mix loves them with and without squeakers. For this dog, it's the best toy we've found.
We can do a little tug of war with it, throw and fetch, and he's even copied various ways we squeak the toy. I don't know what your pup will think, but these seem the most safe, longest-lasting, most engaging toy he has. If we step on it by accident, he always comes running for some fun.
Let us know if you have come up with a new game! We call one "Puppy Puzzle" where we bury it under various objects such as blankets, boxes, and find ways to squeak it when he can't quite find it. It's a daily ritual. We also like to be creative when we introduce new ones by hiding them, or mixing them in with older ones.
For us, it is a family interactive toy. He loses interest faster when playing alone. Plus, because of its size and random bouncing directions, it often gets out of pup's reach under furniture or in spaces too small for him to get it himself. We like playing with our dog, so it works for us. We've tried other toys with similar sizes and appearances, but for whatever reason, this is the one that has become part of daily life, and there appears to be no boredom yet after 2 years. Love!
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Reviewed in the United States on February 1, 2020
★★★★★ 3
Dog won’t bite on these
Size: Small
Great squeak, squishiness, and colors.
Dog won’t bite on latex…
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Reviewed in the United States on March 28, 2026
★★★★★ 5
Good For Heavy Chewers So Far
Color: Onyx Black - Most Durable
Solid quality. My Doberman is a power chewer and I am looking for a toy that he can't destroy in 2 seconds. If a ball lasts more than an hour it is a miracle. It has a sturdy design and is more heavy weight than the other "non-destructible" balls he has. He has only had his ball for one day so we will see how long this ball lasts him. It is the size of a tennis ball, firm, little to no bounce, and seems built for heavy chewing.
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Reviewed in the United States on January 30, 2026
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