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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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4.1 ★★★★★
Based on 12 reviews
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Product Reviews
★★★★★ 2
Not as durable as it looks
Style: Snake
Our half pittbull and half american bulldog tore this apart in 5min.
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Reviewed in the United States on April 27, 2026
★★★★★ 5
Good looking toy that has held up well
Style: Coyote, Style: Coyote
Very durable toy that our little dog especially loves. It's the perfect size for a small dog and she doesn't destroy toys anyway so it's been great. It does feel very durable though and would take our chewer a little while to get to it as well.
I'm not sure it looks like a coyote but the dogs don't seem to mind. It's an attractive toy that looks great and is constructed well so it's not like a slobbery mess when left out.
Good value for the money and would make a great gift because it looks higher quality than most toys.
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Reviewed in the United States on April 14, 2026
★★★★★ 1
May be ballistic nylon, but the seams are flimsy.
Style: Gator, Style: Gator
I really wanted my pup to like this toy because it looks great and feels solid at first. The design is fun, the material seems tough, and it definitely gives the impression that it can handle some rough play. Unfortunately, that didn’t turn out to be the case at all.
After just about 5 minutes of play, the top seam split open and stuffing started coming out. This wasn’t even extreme chewing, just normal play, and it failed almost immediately. As you can see in the photos, the tear happened right along the top seam, which is exactly where you’d expect a toy to be reinforced. Once it opened up, the toy was essentially done.
For something marketed as ballistic nylon, this was really disappointing. It might be fine for very gentle dogs, but if your dog has even moderate chewing habits, this likely won’t last. Based on my experience, I wouldn’t recommend it. especially at 19.99 which is the price at time of my review, I feel it does not offer good value for money.
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Reviewed in the United States on April 9, 2026
★★★★★ 5
A Surprisingly Tough Favorite for All Four of My Dogs
Style: Armadillo, Style: Armadillo
I wasn’t sure what to expect from the Hound 2.0 stuffed armadillo toy, but it has been a total hit in my house. Neither the dogs nor I have managed to find the squeaker yet—so it’s either missing or extremely well hidden—but honestly, the dogs don’t seem to care at all. They love this thing.
I have four dogs ranging from a 22 lb Boston Terrier to an 85 lb pitbull, plus two medium Blue Heelers in between, and this toy has been thoroughly tested by every size and play style. I was a little unsure whether they’d like the harder “shell” portion of the armadillo, but they actually love it. It gives the toy a unique texture and makes it feel more durable.
Before this, I knew nothing about the Hound 2.0 brand, but I’m a fan now. My little Boston Terrier is notorious for ripping every toy he touches, and this armadillo has stood up to him like a champ. The tag said it was made for outdoor play, wrestling, and chewing—and that’s absolutely true. I haven’t taken it outside yet, but inside it has survived rough play from all four dogs and still looks brand new.
I also love that each toy in this line is a different animal with the hard parts in different places. It makes them feel unique instead of just the same toy in different shapes.
I’ll definitely be buying more of these. If you have a pup—big, small, or anywhere in between—I’d recommend giving this toy a try. It’s been a winner in my multi‑dog home.
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Reviewed in the United States on March 10, 2026
★★★★★ 4
Unique armadillo design
Style: Armadillo
I love the look of this armadillo toy. The flexible rubber shell gives it a spiky texture. It has reinforced nylon seams and printed eyes. The feet allow the toy to stand on its own, daring dogs to come knock it over. With a body length of 12" and standing 5" tall, the toy is of good size for even larger dogs, but it intimidated my 8lb yorkie mix. She found the hard shell and spikes off-putting and not fun. The toy is supposed to squeak, but either mine was broken or buried too deep to be useful. This was disappointing because I think a squeak would have helped my pup engage with the toy because she can't resist a noisy toy. I'd recommend this armadillo for larger dogs than my tiny pup. It seems durable with that rubber shell around the body.
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Reviewed in the United States on March 16, 2026