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Description
Mouse DHH 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 preparation and requirements: 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 and
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 preparation and requirements: 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 and mince the tissue. 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. 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 collect the supernatant for analysis. Cell Lysis Buffer: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended 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 freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. 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 utilizes 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 Hedgehog Homolog, Desert (DHH) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of Hedgehog Homolog, Desert (DHH) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Hedgehog Homolog, Desert ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Desert hedgehog, also known as desert hedgehog homolog or Dhh, is a protein encoded by the DHH gene and a member of the hedgehog signaling pathway. The protein encoded by this gene is involved in cell signaling. Several mammalian variants of the Drosophila hedgehog gene (which was the first to be named) have been named after various hedgehog species. The name Desert Hedgehog is in honor of this hedgehog. This gene is not unique to desert hedgehog. It encodes a member of the hedgehog family. The hedgehog gene family encodes signaling molecules that play an important role in regulating morphogenesis. The protein is predicted to be a precursor that is autocatalytically cleaved. The N-terminal portion is soluble and contains signaling activity, while the C-terminal portion is involved in precursor processing. Importantly, the C-terminal product covalently links a cholesterol group to the N-terminal product, confining the N-terminal product to the cell surface and preventing it from freely diffusing throughout the organism. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, and other biological fluids |
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★★★★★ 5
Very happy with purchase
Color: White
This air purifier is great , easy to set up and getting it started , i like that it tells me the air quality and also how quiet it is when running. The timer n the auto settings come in handy. It gets the air clean super fast, definitely worth the price !:)
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Reviewed in the United States on May 8, 2026
★★★★★ 5
Outstanding filter!
Color: White
This has been brilliant at getting dust removed from the air my breathing at night is so much better because it gets pollen out of the air and keeps everything fresh! I haven't used the auto control or sleep mode on purpose but it looks great runs quiet and does a wacking great job!
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Reviewed in the United States on May 2, 2026
★★★★★ 5
Great working air purifier for bedroom
Color: White
Works really well. The pre filter captures so much dust you can really tell it's working as it should. The screen shows how dirty the air is and you can watch it go down as it cleans, didn't think we would need that but it's been surprisingly satisfying to see the quality visually. Very quiet on the lowest setting, easy to sleep with.. gives a nice white noise. The highest still isn't bad, the volume is similar to like a normal tower fan. Design is nice, it's a good looking machine. The only negative would be how bright the small display screen is at night but we just cover it if needed. Haven't tried the oil extract diffuser and probably won't. Works very well for cigarette smell! I'm sure if we had two it would completely remove the smoke smell. Overall would definitely buy again.
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Reviewed in the United States on May 28, 2026
★★★★★ 4
Sound not an issue
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I've had this for about a month, so far works great. I smoke Cigars in my apartment and it is working good. Clears the smoke great even with the filter it came within is a bit loud on the 2nd and third setting, but it is not distracting. I leave it on the first setting over night and its quiet. Next morning there is no cigar smell. Of course I empty ash try. When I come home from work my place smells great and I hardly see see any dust on my glass TV stand and tables. I've read some bad reviews on the sound, but its something you don't hear when tv or radio is on. It really sounds like having a fan on. I would recommend this for the price. I've had other air purifiers no difference on sound.
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Reviewed in the United States on May 9, 2026
★★★★★ 5
MOOKA Air Purifier Rocks!
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I put my Mooka Air Purifier through a very hard test running it 24 hours a day 7 days a week 30 days in a 275 sq ft area so I could provide an accurate review. My husband and I have a dog, woodburning stove, our 115-year-old is close to the road and there is a limestone quarry across the street; our house is extraordinarily dusty. We noticed an improvement the very first day using our Mooka! The nighttime setting is silent so I was able to use it all night. When we’re in the room, we use settings 1 or 2 and can watch TV. When we are out of the room, we use setting 3. Setting 3 can be loud but that is okay, I need that extra power and it does a fantastic job. My Mooka is lightweight, easy to use and a compact workhorse. Another fantastic feature is the highly effective, affordable replacement Hepa filters that have a washable foam pre-filter. I paid under $80 for our Mooka and it is worth every penny. We are ordering a second one. The proof is in our air quality at home. Tried to upload photos of the filter, but could not, sorry!
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Reviewed in the United States on March 15, 2026