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Description
Human PARK2 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 a Parkinson's Disease Protein 2 (PARK2) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of Parkinson's Disease Protein 2 (PARK2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Parkinson Disease Protein 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Parkinson's disease protein 2, also known as Parkin, is a 465-amino acid E3 ubiquitin ligase encoded by the PARK2 gene. Parkin plays a key role in ubiquitination—the process by which molecules are covalently labeled with ubiquitin (Ub) and targeted for degradation by proteasomes or lysosomes. Upon cell injury, Parkin recognizes proteins on the mitochondrial outer membrane and mediates the clearance of damaged mitochondria through autophagy and the proteasome. Parkin also enhances cell survival by inhibiting both mitochondria-dependent and -independent apoptosis. Mutations are associated with mitochondrial dysfunction, leading to the abnormal metabolism that contributes to neuronal death and tumorigenesis in Parkinson's disease. This protein is a component of the multiprotein E3 ubiquitin ligase complex, which in turn is part of the ubiquitin-proteasome system, which mediates targeted protein degradation. Mutations in this gene are known to cause a familial form of Parkinson's disease, autosomal recessive juvenile Parkinson's disease (AR-JP). Furthermore, Parkinson's disease has been described as requiring mitophagy (autophagy of mitochondria). | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, and other biological fluids |
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4.8 ★★★★★
Based on 18 reviews
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Product Reviews
★★★★★ 5
Perfect beginner set
Color: Lightgreen
Love this!!! The toothbrush heads are a little flimsy but overall really liked this for my 3 year old.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 23, 2026
★★★★★ 5
Super Cute, Fun, & Effective
Color: Lightgreen
This set of water flosser and electric toothbrush are just adorable! The shapes of them fit in the hands really comfortably, but also add to the cuteness as the shapes of the animals!
Yes, this is meant for kids - so the toothbrush heads are small (for little kids), and the water flosser doesn't spray terribly hard. BUT they're still very effective.
I especially like the water flosser. It can be taken into the bathtub/shower while bathing, so the spray doesn't end up going places we don't want water to go. It has a small tank of water, and I recommend using room temp water (rather than cold) so that it's comfortable on the gums and teeth. Also to note, the flosser doesn't work when it's sideways or upside-down - it needs to be held upright.
This set is so cute, easy to use, easy to re-charge, and very comfortable. It's also very good quality - a great purchase. I love it!
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Reviewed in the United States on March 17, 2026
★★★★★ 4
4 year old approved!
Color: Lightgreen, Color: Lightgreen
Love that this is a complete set, down to the wall block for the charger!
My daughter is 4 and absolutely loves this set. I personally feel like the toothbrush head is a tad small, but it does do the job well. Its a soft brush head, with plenty of replacement heads. The toothbrush and the flosser both arrived fully charged, and so far we havent had to recharge it, but we're only a few days in. So, it seems to have a pretty good battery life so far. (I will update my review if anything changes on that aspect)
The toothbrush and flosser both stand upright independently on the counter, without tipping over.
The water flosser is hard fir my daughter to do by herself. She sprays water everywhere, as theres several buttons on it and she cant seem to remember in time which button turns it off. I help her with flossing and its been going fine.
Whether youre on high or low the water pressure is not too hard for a kids mouth. In fact, I think the low setting does nothing but rinse the mouth and basically teach the habit of flossing. We use the high setting and it does and OK job. I am a little old school though, and feel like nothing will top a good ole dental floss flossing, but water flossing is a safer route to avoid cutting gums or getting floss stuck between close teeth.
The sound level is minimal, a fraction of the sound level my adult water flosser makes. It doesnt shake/vibrate too badly in the hand and theyre bith comfortable to hold, and size appropriate for my (4yrs) daughters hands.
It comes in a really nice box, packaged perfectly, making this a great gift for your little ones. The colors and theme makes brushing teeth exciting and fun again for your littles :)
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on October 25, 2025
★★★★★ 5
Great for kids
Color: Lightgreen
The HUACOCARE Kids Water Flosser and Electric Toothbrush Combo is an absolute game-changer for establishing a solid dental hygiene routine with children. The light green giraffe and monkey design is incredibly charming and immediately turned what used to be a bedtime struggle into a fun activity that my child actually looks forward to. Having both the sonic toothbrush and the water flosser in one compact unit is brilliant, as it saves counter space while ensuring that every part of their mouth gets a professional-level clean. The three gentle flossing modes and two sonic brushing modes are perfectly calibrated for younger, more sensitive gums, providing a thorough clean without any discomfort.
One of the standout features of this set is how braces-friendly it is; the water flosser reaches those tricky spots around wires and brackets that a standard toothbrush simply can't touch. It is clear that a lot of thought went into the ergonomics and the "fun factor," making it a perfect gift that is both practical and engaging. Since we started using this combo, our dentist has noticed a significant improvement in plaque removal and overall gum health. If you are looking for an all-in-one solution to build healthy brushing habits and make dental care feel like less of a chore, this set is worth every penny.
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Reviewed in the United States on February 21, 2026
★★★★★ 5
They Work!
Size: 60 Count (Pack of 1)
Thanks to my mouthrinse, I think, I have no issues with my teeth, gums, or breath, but cold air does inflame my sinuses, ETs, and ears. And I have to travel to cold places often. I hate that inflammation! As a classical singer, I really hate it! I have to keep my throat, sinuses, ETs and ears healthy. These probiotics have helped me to do just that. Previously, I tried prescription anti-inflammatories, and while they worked, anti-inflammatories can have undesirable side effects. I didn't experience any, but who knows? They could have been brewing. And I don't like chemical drugs. Much better to restore a healthy balance of bacteria in my upper respiratory system. These probiotics are so easy to take. I just let one dissolve in my mouth in the morning and one in the evening. I take them after breakfast, after brushing my teeth, and after brushing my teeth in the evening. It took a few weeks, but then I started seeing results. My sinuses were clearer, my ETs and ears would no longer close against the cold as they did previously. I no longer get the roaring in my right ear that I got when awakening due to sinus drainage. And I'm sure these probiotics are helping my teeth, gums, and breath even though I didn't really have any issues with them previously. If you decide to take these, and I highly recommend you do, be sure to let them dissolve or chew them slowly. To swallow them whole, which can be done easily, defeats their purpose since they are oral probiotics. And remember, these aren't probiotics for gut health. For that, you need a different probiotic mix, with different cultures. I do not refrigerate my probiotics, ever, and they remain potent. The taste is a light mint, and it's very agreeable. Not bad at all. Even if it were, the health benefits would definitely outweigh it. Bottom line: If you're experiencing any issues with your teeth, gums, breath, sinuses, throat, ETs, or ears, then these probiotics are for you! Try them! I think you'll be pleasantly surprised, and greatly pleased, just like I was!
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Reviewed in the United States on January 27, 2020
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