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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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4.4 ★★★★★
Based on 15 reviews
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Product Reviews
★★★★★ 5
my new favorite book
Format: Kindle
Ok so I never write reviews but this book was so good I felt the need to write this. Firstly your introduced to Huntyr you see her closed off hard core badass than towards the end you see the most subtle change and growth it’s amazing and the enemies to friends to lovers was just perfect, AND THE TWIST AT THE END GOT ME GOOD! You see one spicy scene the whole book but it doesn’t even MATTER BECAUSE THE BOOK WAS THAT GOOD. I’ve read 85 books in 2023-2024 so far and I’m pround to say this is my all time favorite. I’m so excited to read more of Emily Blackwoods books, this was my first time reading one of hers and I’m glad I did because HOLY!! Well done Emily well done
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Reviewed in the United States on February 23, 2024
★★★★★ 4
Fast paced romantasy you will not want to put down!
Format: Kindle
4.25 stars! I LOVED this book with similar vibes to Hush Hush, Fourth Wing, and The Serpent and the Wings of Night! It was fast paced with easy world building and will keep you turning the pages late into the night because you will not want to put it down!
Huntyr is a fierce bad@ss FMC trained to kill vampyres her entire life. She is sent on a mission to go to the academy and earn her spot into The Golden City. Upon arrival, she is forced to room with the delicious fallen angel, Wolf, who is the only one who knows about her assassin identity. The romance, the plot twists, the secrets revealed, the battles, and the tantalizing training scenes had me hooked! And that ending…. I’m holding my breath in need to know hell!
Read if you love:
🪽 Fae, Vampyres, Fallen Angels
🪽 Academy setting with magical trials
🪽 Forced proximity and slow burn
🪽 Rivals to lovers
🪽 Hidden identities and secrets
🪽 Tend your wounds
“𝘖𝘧 𝘤𝘰𝘶𝘳𝘴𝘦 𝘐 𝘸𝘢𝘴 𝘸𝘢𝘵𝘤𝘩𝘪𝘯𝘨 𝘺𝘰𝘶. 𝘐 𝘤𝘰𝘶𝘭𝘥𝘯’𝘵 𝘭𝘰𝘰𝘬 𝘢𝘸𝘢𝘺 𝘧𝘰𝘳 𝘢 𝘮𝘰𝘮𝘦𝘯𝘵, 𝘦𝘷𝘦𝘯 𝘪𝘧 𝘐 𝘵𝘳𝘪𝘦𝘥.”
“𝘐𝘧 𝘺𝘰𝘶 𝘸𝘢𝘯𝘵 𝘮𝘦 𝘰𝘯 𝘮𝘺 𝘬𝘯𝘦𝘦𝘴, 𝘏𝘶𝘯𝘵𝘳𝘦𝘴𝘴, 𝘢𝘭𝘭 𝘺𝘰𝘶 𝘩𝘢𝘷𝘦 𝘵𝘰 𝘥𝘰 𝘪𝘴 𝘢𝘴𝘬.”
“𝘠𝘰𝘶 𝘥𝘰 𝘯𝘰𝘵 𝘬𝘯𝘰𝘸 𝘵𝘩𝘦 𝘷𝘪𝘰𝘭𝘦𝘯𝘤𝘦 𝘵𝘩𝘢𝘵 𝘳𝘶𝘯𝘴 𝘵𝘩𝘳𝘰𝘶𝘨𝘩 𝘮𝘺 𝘷𝘦𝘪𝘯𝘴, 𝘣𝘦𝘨𝘨𝘪𝘯𝘨 𝘮𝘦 𝘵𝘰 𝘰𝘣𝘭𝘪𝘵𝘦𝘳𝘢𝘵𝘦 𝘢𝘯𝘺𝘰𝘯𝘦 𝘸𝘩𝘰 𝘭𝘢𝘺𝘴 𝘢 𝘩𝘢𝘯𝘥 𝘰𝘯 𝘺𝘰𝘶.”
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Reviewed in the United States on June 12, 2024
★★★★★ 5
Excellent Rivals to Lovers!!
Format: Kindle
The tension and banter between Huntyr and Wold was delectable. I absolutely love the fallen angel and all of his flaws. Huntyr is amazing too being a badass FMC with some major trauma. The world building was great and I enjoyed the training aspect of the story. The writing was immersive and was in the story the whole time. The ending had quite a twist that I hadn’t anticipated and made my jaw DROP. Excellent job! I also loved the narration. Laura is one of my fave narrators!
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Reviewed in the United States on August 15, 2025
★★★★★ 5
Fallen Angels, fae, vampires, oh my!
Format: Kindle
Rating: 4.5 | Spice: 2 (but a good slow-burn)
•
Main Characters: Huntyr and Wolf
•
I couldn’t wait to read this book; there was so much hype about it! And there was no doubt why. I fell in love with the characters and the plot itself. This book is mainly plot driven more than friction driven but it’s easy to follow along with. The characters are fun, easily understood. The main setting is at an academy where both the main characters are going through trials and building strength for the final test, The Transcendent. There are fantastic side characters as well. I loved the camaraderie between Huntyr and her friends. But we don’t like Lanson. 😆
We do have some plot twists that come into play throughout the book. Secrets and betrayal to be seen. I did adore Wolf and Huntyr’s relationship. It was a classic slow burn trope. They didn’t hit it off fast, but in time their feelings grew. I loved their banter, so sexy. Wolf is your next book boyfriend; Huntyr is your next vampire assassin independent bad-a*s female. Themes include loyalty, trust, self-discovery, a true slow burn romance. Side note: book ends on a angsty cliffhanger!
•
Emily, thank you for writing this awesome novel and I cannot wait to devour Book 2, Blood So Brutal! 😍
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Happy reading, my lovelies! xo
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Reviewed in the United States on January 21, 2024
★★★★★ 4
“My heart bows to you and you only, Huntress.”
Format: Kindle
3.5 🌟
This book popped up in my KU recommended reading suggestions and the synopsis sounded like what I was in the mood for. I'm so glad I took a chance on it. I went into this knowing absolutely nothing about it and ended up really liking it. I love when this happens.
The main characters are likeable and I easily found myself rooting for them. There is a mystery element to each of their backstories that I enjoyed watching unfold and can't wait to get more of. Wolf, in particular, has me fixated. Love him.
I found this to be an entertaining, addictive read with a plot that moves along at a good pace. It reads so easily I found myself very reluctant to put it down. Lots of twists and turns and the angst is there. A good set up for the next book to come, for sure.
My issues with this book....the dialogue feels a bit juvenile at times and there is a repetitive over use of a particular word phrasing that I found myself giving the ole eye-roll to. There are, without a doubt, some pretty cliche moments that gave me a bit of the cringe. I think this could've certainly 100% benefited from more depth regarding the world building. Perhaps the world building was sacrificed to keep the pacing quick? Just a guess. Also, the lack of consistency of character for the FMC was really evident and so she feels quite illogical at times.
Overall, this was a fun and enjoyable read that hit the spot well enough for me. That ending certainly has me impatiently pining for book 2!
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Reviewed in the United States on June 18, 2024