Pay in installments of $46.03 with
,
and
Shipping Estimate
USA
- USA
- CAN
- USA
- CAN
Ships within 48 hours · Estimated delivery Aug 11 - Aug 16
Promo Codes Available:
For Your Every Summer RSVP, with Code: SUMMER15
Description
Human PP 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. |
|||||||||||||||||||||||||||||||||
| 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 Parkin protein (PP) capture antibodies. 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 Parkin protein (PP) content in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Parkin proteinELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
|
|||||||||||||||||||||||||||||||||
| Background | Parkin (PP) is a protein encoded by the PARK2 gene. It is a component of a multiprotein E3 ubiquitin ligase complex, which in turn is part of the ubiquitin-proteasome system, which mediates targeted protein degradation. Mutations in this gene cause a familial form of Parkinson's disease, autosomal recessive juvenile Parkinson's disease (AR-JP). Furthermore, Parkin has been described as essential for mitophagy (autophagy of mitochondria). Furthermore, Parkin contains a C-terminal motif that binds to PDZ domains. It has been shown to bind to the PDZ domain-containing proteins CASK and PICK1 in a PDZ-dependent manner. | |||||||||||||||||||||||||||||||||
| 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. |
|||||||||||||||||||||||||||||||||
| 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 |
Shipping Notes
- Free Standard Shipping on $100+ Orders to the USA.
- Except Preorder products are shipped in 48 hours.
- Delivery to the USA:
- Standard Shipping : 3-10 business days
- If time is of the essence, please consider selecting expedited delivery for faster service.
Exchange/Return Notes
- We offer a 30-day return/exchange service after receiving.
- Final sale items are not eligible for returns or exchanges.
- To process your return/exchange, please contact us at [email protected]
- Please click here for more details>>> Return & Exchange Policy
4.5 ★★★★★
Based on 21 reviews
Sort
Product Reviews
★★★★★ 4
Good Coverage But...
Asurion had me to package up my Ipad and send to them to research the problem with it overheating and cracking the screen near the home button. The turnaround was about 6 days from when i shipped it next day and them returning the device. When I began to setup the IPad the first thing I notice was an error message stating touch id could not be setup on the device (it worked previously) and the ipad still got hot when charging. I had to return it to them a second time to "resolve" the issue. They determined they couldn't fix the issue and returned the cost of the device. The only problem is you can't replace a device for what it cost at that time of purchase, especially now when prices have soared. They need to fix or replace the device at the cost of what iit costs at the time of replacement.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 8, 2023
★★★★★ 5
Must buy for your tablet!
Amazing service, cannot recommend enough! So easy to start a claim, and super fast turn around time, I was informed through every step.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 9, 2025
★★★★★ 5
They sent me a Amazon gift card
I insured an iPad for my son and had it repaired multiple times from him stepping on it. Quick and fast repairs. The last time he accidentally damaged it they sent me an Amazon gift card for the full price I originally paid for it. I was able to use the gift card right away because it was digital. I used it on an iPad that was on sale for half the original price so I was able to get him the night light he always wanted.
Best service available and I insured this iPad right away with them.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 10, 2023
★★★★★ 1
They will do anything to get out of fixing or replacing your item.
I bought a refurbished Samsung Galaxy Tablet S6 from Amazon through Affirm. I've already had the tablet for nearly 3 months. The tablet is defective. After barely 3 months it won't charge. I attempted to file a claim. The Asurion website tells me to return the item. Samsung doesn't make these tablets anymore. You cannot buy a brand new tablet of this model. They aren't sold online. The reason I bought the warranty which was over $100 is so that Asurion will fix the tablet if there is a problem. They were unwilling to make any effort to repair or fix this rare tablet. The website says to return the item. This is a lengthy return process. Amazon will not give me a QR code or let me print a label that I can take to a local UPS store. The only place Amazon will let me drop off the item is 2 or 3 towns away. The only option Amazon is giving me is to have UPS come pick up the tablet without giving me a label. I had to purchase a box from UPS and have them package it for me without providing me a label because Amazon says I don't need one. On the first attempt the UPS guy wasn't able to pickup the item because they need a special kind of form and my UPS driver didn't have the form. I have to wait until a UPS driver with the proper form is able to pick up my item before they can start the return process. Now I can't buy a Samsung Galaxy Tablet S6 from Amazon unless I want to go through this or a similar process every time an Amazon 3rd party seller sells me a defective product. Thanks for nothing Asurion. I'm going to start buying my electronics in person from Best Buy. They sell electronics that work and they immediately replace damaged and defective items that are under warranty. Amazon should ban Asurion from selling warranties on their website. They are selling warranties that they have no intention of providing the end product for. This is the second I've had a problem with Asurion. The first time I had a problem with a pair of headphones purchased through Amazon that were covered by my Asurion warranty that I had purchased. Asurion wouldn't help me and directed me to try and use the manufacturer warranty. When I buy a warranty from someone it's because I want the seller of the warranty to solve my problem. Not tell me to return the item or direct me to use the manufacturer warranty. It's not my fault the refurbisher couldn't make the tablet function properly for longer than 3 months.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on October 14, 2022
★★★★★ 5
Apple is number one
Style: Wi-Fi, Size: 128GB, Color: Blue, Configuration: Without AppleCare+
The best iPad it’s worth the money it works perfectly fine I think the battery life is fine, but it probably needs a little bit of work, I would choose Apple anytime of the week. The durability is perfect and everything and the gameplay is absolute amazing. this is also for personal use and probably college use. I’m probably going to use this for college as well.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 5, 2026