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Description
Mouse ALDH2 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 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 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. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 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 competitive enzyme-linked immunosorbent assay (ELISA). Samples, standards, biotin-labeled antibodies, and HRP enzyme conjugates are added sequentially to microwells pre-coated with the universal species Aldehyde dehydrogenase, mitochondrial (ALDH2) antigen. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue under the catalysis of peroxidase (HRP) and to the final yellow under the action of acid. The intensity of the color is negatively correlated with the universal species Aldehyde dehydrogenase, mitochondrial (ALDH2) in the sample. The absorbance (OD value) is measured at a wavelength of 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Aldehyde dehydrogenase, mitochondrial ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Aldehyde dehydrogenase is an enzyme encoded by the ALDH2 gene. This protein belongs to the aldehyde dehydrogenase family of enzymes. It is the second enzyme in the primary oxidative pathway of ethanol metabolism. Two major hepatic isoforms of ALDH2, cytosolic and mitochondrial, can be distinguished by their electrophoretic mobility, kinetic properties, and subcellular localization. Disorders associated with ALDH2 include alcohol sensitivity, acute alcoholism, and Ahmed syndrome, two genes involved in the gene. Pathways involved include neurotransmitter clearance at the synaptic cleft and biotransformation of metabolic pathways. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.4 ★★★★★
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Product Reviews
★★★★★ 5
From Pixels to Problems! Great read!
Format: Hardcover
“Play Nice” offers an enjoyable deep dive into the tumultuous history of Blizzard Entertainment, chronicling its journey from a ragtag group of brilliant college students to its evolution under corporate ownership and its current state. Schreier provides fascinating insights into the antics of Blizzard’s early employees, showcasing their outlandish attitudes, relentless work ethic, and tight-knit camaraderie.
The book explores how Blizzard transitioned from a company renowned for producing high-quality, polished games that left competitors in the dust to one struggling to preserve its heart and soul amid mounting corporate pressures. While the corporate side and C-suite executives are often cast in a negative light, Schreier thoughtfully examines the motivations behind their decisions, offering perspectives from all levels of the company—from executives and middle management to QA testers. This balanced approach provides a refreshing take, avoiding oversimplified blame and instead considering multiple sides of the story.
And while it’s easy to villainize the suits in the boardroom, Schreier does a great job showing why some decisions were made. From executives to QA testers, he pulls back the curtain to reveal a mess of perspectives, reminding us that every bad decision has some kind of reason behind it (even if it’s still a bad decision).
The book also revisits the scandals that put some serious smudges on Blizzard’s reputation, offering new angles and fresh commentary. As someone who once lived for Blizzard games—cheering at Overwatch League matches and losing entire weekends to Diablo marathons—I can’t help but root for Blizzard to find its way back to glory. And hey, if it means waiting another decade for their next masterpiece, so be it. It’s done when it’s done.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 6, 2025
★★★★★ 5
Reads like your favorite succession episodes
Format: Hardcover
Great book—thoroughly researched and delightfully written! Highly recommend to all my gamers and friends from that era.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 12, 2025
★★★★★ 4
Great insight into an otherwise obscure world
Format: Hardcover
As someone who grew up playing blizzard's games for an unfathomable amount of hours I've always been interested into their inner workings, especially considering their downfall in recent years. This book holds a ton of information and knowledge, is well sourced, and is the work of someone with obvious deep familiarity with the industry and its particularities.
Besides the information itself, the book it written in fun and interesting prose, and it keeps the rhythm fast and entertaining, so it reads more like a novel than a journalistic article.
Overall, an entertaining piece of insight into a world that is normally quite unknown, even to long time gamers like myself.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 15, 2026
★★★★★ 5
Great read
Format: Hardcover
Extremely interesting book
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 3, 2026
★★★★★ 3
Great insight on what happened at Blizzard but...
Format: Kindle
My main issue with the book is the lack of non-american stories that explained the bigger picture. As a former Blizzard dev, there's much more than what happened in Irvine and Korea, with Europe's office mentioned almost as a footnote, and nothing else from the other regional stories. Shame but I guess the book would've been double the size.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 12, 2025
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