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Description
Mouse ALDH 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 specimens using EDTA or heparin as an anticoagulant and centrifuge at 1000×g for 15 minutes at 2-8°C within 30 minutes of collection. Remove the supernatant for testing, or store at -20°C or -80°C, but avoid repeated freezing and thawing. Cell culture supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for testing, or store at -20°C or -80°C, but avoid repeated freezing and thawing. Pre-Test Preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare a gradient standard working solution: Add 1 mL of universal diluent to the lyophilized standard. Let stand for 15 minutes to completely dissolve, then gently mix (concentration 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 500 μL of universal diluent to each tube. 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 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 acetaldehyde dehydrogenase (ALDH) 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 acetaldehyde dehydrogenase (ALDH) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Acetaldehyde dehydrogenase | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Acetaldehyde dehydrogenase (ALDH), a type of aldehyde dehydrogenase, catalyzes the oxidation of acetaldehyde to acetic acid. Alcohol dehydrogenase in the liver oxidizes ethanol (a component of alcohol) to acetaldehyde. The resulting acetaldehyde serves as a substrate for further conversion to harmless acetic acid (a component of vinegar) under the catalysis of ALDH. Aldehyde dehydrogenases are polymorphic enzymes responsible for oxidizing aldehydes to carboxylic acids, which are metabolized by the body's muscles and heart after leaving the liver. These enzymes exist in three distinct classes in mammals: class I (low Km, located in cell membranes), class II (low Km, located in mitochondria), and class III (high Km, such as those expressed in tumors, stomach, and cornea). Within these three classes, there are constitutive and inducible forms. ALDH1 and ALDH2 are the most important aldehyde oxidases, both tetrameric enzymes composed of 54 kDa subunits. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, cell culture supernatant |
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4.5 ★★★★★
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Product Reviews
★★★★★ 5
Quite nice and very functional
Size: 7.7" x 7.7" x 1.7", Color: Grey (Metal Silvered), Size: 7.7" x 7.7" x 1.7", Color: Grey (Metal Silvered)
Elegant and substantial - this valet catch all for your night stand or dresser is quite nice and very sophisticated. Sleek and soft brushed suede like material. It was packed well, and is quite nice to keep miscellaneous items corralled together. I’m very pleased and would recommend.
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Reviewed in the United States on February 15, 2025
★★★★★ 5
Perfect size, high quality
Size: 7.7" x 7.7" x 1.7", Color: Black (Metal Silvered)
I had a previous catch all tray that I bought on amazon, it used buttons on the 4 corners to turn a flat cardboard into a tray. Over time, the catch all tray got filled up, not just with a wallet, keys and eyeglasses but eventually business cards and other knick knacks. With all the items inside the tray, sometimes the buttons would start to pop out. It wasn't awful, more of an annoyance. But it was time for me to find a more durable tray. I looked on amazon and added 6 different trays that had "potential" into my shopping cart. Then I carefully evaluated each one and deleted them out of my cart until there was only one left. Some had multiple compartments. Some were larger than this one. I felt the ones with compartments would limit my ability to put the things I want in there. Also, I didn't want it so big that it took up too much desk space. Ultimately, I felt this was the perfect size. I'm extremely happy with it, the quality is really nice and it looks and feels durable. No more loose buttons and spilled knick knacks.
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Reviewed in the United States on October 14, 2024
★★★★★ 5
Keeps clutter away
Size: 7.7" x 7.7" x 1.7", Color: Black (Metal Silvered)
This works great to corral all my notes I write on post its or small notes. Keeping them hidden from the desk top b/c or the depth but still visible to me. Much better than keeping on the plotter. Looks great in quality .
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Reviewed in the United States on October 25, 2025
★★★★★ 5
Attractive, Simple, And Well-Made
Size: 7.7" x 7.7" x 1.7", Color: Orange (Metal Glided)
I use this as a "catchall". A place to stash things like my wallet and keys when I enter the door, so that I don't use them (I had been prone to that!). It serves it's purpose well. It's both sturdy and attractive, and a good size for a "catchall". Recommended for those like me who need such things.
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Reviewed in the United States on April 8, 2024
★★★★★ 4
Good quality, orange color
Size: 7.7" x 7.7" x 1.7", Color: Orange (Metal Glided), Size: 7.7" x 7.7" x 1.7", Color: Orange (Metal Glided)
The quality is surprisingly good but the color is much more orange toned in person.
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Reviewed in the United States on April 17, 2024
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