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Description
Human ADA 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: 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: 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. 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. 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. 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 an adenosine deaminase (ADA) capture antibody. 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 adenosine deaminase (ADA) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Adenosine Deaminase ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Adenosine deaminase (also known as adenosine aminohydrolase, or ADA) is an enzyme involved in purine metabolism. It breaks down adenosine from dietary sources and utilizes it for nucleic acid turnover in tissues. Its primary function is the development and maintenance of the immune system. However, the full physiological role of ADA is not fully understood. ADA exists in both small (monomer) and large (dimer) forms. In the monomeric form, the enzyme is a single polypeptide chain folded into eight parallel α/β barrels surrounding a deep central pocket, which serves as the active site. | |||||||||||||||||||||||||||||||||
| 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 and other biological fluids |
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4.1 ★★★★★
Based on 25 reviews
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Product Reviews
★★★★★ 5
Emotional read with children
Format: Kindle
Cammie is a single mother of two children whose ex-boyfriend, who is an alpha that he met his scent matched omega and dumped her via messenger on his phone then the next day put their house up for sale. Cammie moves back to her hometown, but because she has no support, is working two jobs to keep her and the children in a small rented house. One night her four year gets out of the house and she meets a neighbour, but she refuses to make the same mistake again by getting involved with another alpha, but Reid is convinced they are scent matched mates even though Cammie is a beta. Reid and his omega Fin start to court Cammie and her two children that they belong together as a pack and family. Cammie's ten year old son is a tough one to convince, but the scenes between him and Reid and Fin, they were so good. I was tearing up at times, found family was a big theme and I really liked this book. I highly recommend that you try it too.
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Reviewed in the United States on February 16, 2025
★★★★★ 5
Wonderful
Format: Kindle
What a wonderful long read, with depth, emotions, storyline and a touch of $mut. This story was definitely more centered around the storyline rather than any $ex scenes and lived up to “Cozy Omegaverse”. We saw the pack dynamic’s evolve as she was ready for them to evolve, they were patient with her and allowed her to break free from her own constraints.
Amazing!
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Reviewed in the United States on June 25, 2025
★★★★★ 4
A Nice Story
Format: Kindle
I love Omegaverse and its many stories. The book is a wonderful story. The characters are intriguing and are each unique. There were a few issues with repetitive phrases through out the book and a few misspellings but the biggest issue was the very painful slow burn. I feel if it weren’t so slow
It could have incorporated more content about each character. Overall a solid story though.
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Reviewed in the United States on April 25, 2025
★★★★★ 3
decent…but
Format: Kindle
This is my first book from this author, and while I do really love omegaverse stories, this one left a lot to be desired. This book is a hard-core slow, slow, slow burn: (75% or later in the book)
I will say that I finished it. Unfortunately, I struggled through a lot of it due to punctuation errors, and general editing that must have been looked over during the revision process.
I also feel like a lot of information was left out and there wasn’t closure regarding her parents. The dynamic of the pack before Lydia joined is a little confusing, the only person who showed affection towards Elias was the Head alpha. The other two seemed indifferent to him in an intimate relationship way. That doesn’t really scream cohesive pack to me.
I am hopeful that the next book in the series is better edited, and that the author decides not to use the same phrases over and over again. I’m pretty sure “honey-warm” was mentioned over 15 times when describing Elias’s scent. That got really old, really fast.
Overall, it was a decent book. Not sure I would read it again or recommend it until of these errors are remedied.
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Reviewed in the United States on April 26, 2025
★★★★★ 5
Definitely a comfort read
Format: Kindle
Just a comfy cozy nice long read. While this is definitely a SLOOOOOOOOWWWWW burn, this is how you do RH right; a nice long novel where you actually get to know the characters so you can feel more connected to them & the story, not to mention getting enough time to actually remember all their names haha. & they are all consent kings. Also I do so love me some double omega OV, siiiigh (heart eyes). I wish there had been juuust a bit more spice, like maybe just between Elias & others before MC was ready (made Elias feel a bit underdone or gimmicky without it), I did love this read. Low stress, comfy read. It could've used another round of editing for some minor errors & type issues but it wasn't bad enough to pull ne out of the story. Though I did giggle at 'knecks', thinking kuh-necks in my head. Overall I'd say 9/10
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Reviewed in the United States on March 28, 2025