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Description
Human DDC 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 handling and requirements: The detection range of the kit is not equivalent to the concentration range of the analyte in the sample. Before the experiment, it is recommended to estimate the analyte concentration in the
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 handling and requirements: The detection range of the kit is not equivalent to the concentration range of the analyte in the sample. Before the experiment, it is recommended to estimate the analyte concentration in the sample based on relevant literature and conduct preliminary experiments to determine the actual concentration in the sample. If the analyte concentration in the sample is too high or too low, dilute or concentrate the sample appropriately. If the sample type is not listed in the instructions, it is recommended to conduct a preliminary experiment to verify the validity of the test. Serum: Collect whole blood in a serum separator tube and place it at room temperature for 2 hours or at 2-8°C overnight. Then centrifuge at 1000×g for 20 minutes and remove the supernatant. Alternatively, store the supernatant at -20°C or -80°C, but avoid repeated freezing and thawing. Plasma: Collect the sample using EDTA or heparin as an anticoagulant. Within 30 minutes of collection, centrifuge at 1000×g for 15 minutes at 2-8°C. Remove the supernatant and test it. Alternatively, store the supernatant at -20°C or -80°C, but avoid repeated freezing and thawing. Tissue homogenate: Rinse the tissue with pre-chilled PBS (0.01M, pH 7.4) to remove any residual blood (lysed red blood cells in the homogenate may affect the test 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., 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 or in a homogenizer. To further lyse tissue cells, the homogenate can be sonicated or repeatedly freeze-thawed. Finally, centrifuge the homogenate at 5000×g for 5-10 minutes and remove the supernatant for analysis. Cell culture supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant and analyze it immediately, or store it at -20°C or -80°C, but avoid repeated freeze-thaw cycles. Cell lysis buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and harvest the cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Wash the collected cells three times with pre-chilled PBS and resuspend in 150-200 μL of PBS per 1 × 10^6 cells (protease inhibitors are recommended; if the cell count is very low, the PBS volume can be reduced 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 testing. Other biological samples: Centrifuge at 1000 × g for 20 minutes, and remove the supernatant for testing. Sample Appearance: The sample should be clear and transparent, and any suspended matter should be removed by centrifugation. Sample Storage: Samples collected for testing within one week can be stored at 4°C. If testing is not possible, aliquot the sample into single-use aliquots and freeze at -20°C (for testing within one month) or -80°C (for testing within six months). Avoid repeated freeze-thaw cycles. Hemolysis of the sample can affect the final test results, so hemolyzed samples are not suitable for this test. Pre-Assay 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 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: Add 500 μL of universal diluent to each of seven EP tubes. Pipette 500 μL of the 10 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5 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, as shown in the figure below. 3. Prepare HRP antibody working solution: 15 minutes before use, centrifuge 100 μg of concentrated HRP antibody at 1000 × g for 1 minute. Dilute the 100 μg of concentrated HRP antibody to a 1 μg working concentration with universal diluent (e.g., 10 μL of concentrated solution + 990 μL of universal diluent). Prepare immediately before use. 4. Preparation of 1× Wash Buffer: Dissolve 10 mL of 20× Wash Buffer in 190 mL of distilled water (Concentrated Wash Buffer removed from the refrigerator may crystallize, which is normal. Allow to stand at room temperature until the crystals have completely dissolved before preparing the buffer). 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 them 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 sealant and incubate at 37°C for 60 minutes. (Recommendation: Dilute the samples to be tested at least 1-fold with Universal Diluent before adding them to the plate. This minimizes the impact of matrix effects on the test results. When calculating sample concentrations, multiply by the corresponding dilution factor. It is recommended to run replicates for all samples and standards.) 3. Wash: Discard the liquid and add 300 μL of 1x wash buffer to each well. Let stand for 1 minute, shake off the wash buffer, and pat dry on absorbent paper. Repeat this process three times (a microplate washer can also be used). 4. Add HRP detection antibody: After washing, add 100 μL of HRP detection antibody working solution directly to each well. Cover with a film sealer and incubate at 37°C for 60 minutes. 5. Wash: Discard the liquid and wash the plate five times according to the procedure in step 3. 6. Add substrate: Add 90 μL of substrate (TMB) to each well, cover with a film sealer, and incubate at 37°C in the dark for 15 minutes. 7. Add 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 450 nm. Calculation of Experimental Results: Result Interpretation: 1. Calculate the average OD value of the standard and sample replicates and subtract the OD value of the blank well as the correction value. 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 it appropriately and retest. Multiply the sample concentration by the corresponding dilution factor when calculating the sample concentration. |
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| Sensitivity | 0.06 ng/mL | ||||||||||||||||||||||||||||||
| Theory | This kit uses a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, and HRP-labeled detection antibody are sequentially added to microwells pre-coated with human DOPA decarboxylase (DDC) capture antibodies. After incubation and washing, the sample is developed with the substrate TMB. TMB converts to blue under the catalysis of HRP peroxidase and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of human DOPA decarboxylase (DDC) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | ||||||||||||||||||||||||||||||
| Source | Human | ||||||||||||||||||||||||||||||
| Synonym | Human Dopa Decarboxylase ELISA Kit | ||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | ||||||||||||||||||||||||||||||
| Composition |
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| Background | Aromatic L-amino acid decarboxylase (AADC or AAAD), also known as dopa decarboxylase (DDC), tryptophan decarboxylase, and 5-hydroxytryptophan decarboxylase, is a lytic enzyme. This enzyme uses pyridoxine phosphate (PLP), the active form of vitamin B6, as a cofactor. PLP is essential for the decarboxylation mechanism of AADC. In the active enzyme, PLP binds to lysine-303 of AADC as a Schiff base. | ||||||||||||||||||||||||||||||
| 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 | 0.15-10 ng/mL | ||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates, cell lysates and other biological fluids |
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4.1 ★★★★★
Based on 20 reviews
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Product Reviews
★★★★★ 5
Phoebe approved! 3 months and still going!!
Color: Hammer, Size: Large, Color: Hammer, Size: Large
I have 2 dogs--a chocolate lab and a pittie mix. Normally, they shred their toys within minutes/hours if not days, but this one has lasted for over 3 months! It is well loved and starting to show some wear, but I am super impressed with how long this one has lasted. It has a couple holes in it but they haven't disemboweled it. The squeeker didn't last too long, but they never do.
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Reviewed in the United States on April 7, 2026
★★★★★ 3
Good for modest chewers who don't destroy toys.
Color: Hammer, Size: Large
We have four dogs, one of whom is a super chewer. Unfortunately, this toy wasn't even durable enough to stand up to our mid-level chewer. She had the squeaker out of this in mere moments, and the rope was untied and in shreds. We had to basically remove the squeaker from her, then clip the rope from the toy so she didn't try to eat it. Now, don't get me wrong. She's still having a great time with this toy, even as she whips it around and tosses stuffing everywhere. This is definitely sized for medium to large dogs, but I would only recommend this to dogs who don't like to destroy their toys, but instead just like to squeak them and maybe play some light tug. For those dogs, this will be a great option. For super chewers, this will last only minutes and I would not recommend it.
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Reviewed in the United States on March 24, 2026
★★★★★ 5
Fun Toy for Fun Times
Color: Hammer, Size: Large
Perfect fun toy for my dog!
I bought this squeaky plush toy for my dog and it has been a huge hit! The design is adorable — like a little toolbox toy — and the material feels soft yet durable enough to handle lots of chewing. The squeaker sound gets my dog’s attention every time and keeps him entertained for ages. It’s a great size for playtime and fetch, and so far it’s holding up really well against rough play. Highly recommend this if you’re looking for a fun and engaging toy for your furry friend!
She chews it up and has a good time. Good price.
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Reviewed in the United States on February 8, 2026
★★★★★ 5
Very durable for a plush toy
Color: Hammer, Size: Large
My GSD husky mix loves this toy. She is "working on it" as I write. Her first call to action with any new toy is to get the squeaker out. This toy is well made enough so that she has not been able to reverse engineer it immediately (as she does with cheap dog toys). She seems to enjoy chewing on the handle making it squeak, which is pretty cute. The toy itself is bigger than I thought it would be which is nice, as my dog is bigger too. It is soft and well stuffed. The rope on the end will make it a great tug toy too. Right now it has my dog's full attention so we shall see how long it lasts. Overall, while not indestructible, it's definitely better than 90% of the toys we've gotten her so far in terms of durability. Great product!
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Reviewed in the United States on January 22, 2026
★★★★★ 4
Seems sturdy so far
Color: Screwdriver, Size: Large
So far my boys haven't ripped this apart, which is a good sign. They like to play tug with it and the squeaker is engaging (my pittie loves squeakers). Here's hoping it continues to last with my 2 tough chewers.
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Reviewed in the United States on December 31, 2025