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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. |
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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 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 |
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| 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. |
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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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4.6 ★★★★★
Based on 20 reviews
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Product Reviews
★★★★★ 5
It smells as good as my cologne.
I never go out without a shot of Guerlain L'Homme Idéal L'Intense cologne. Guerlain does not offer a layered line of products with similar scents. I wanted a deodorant and body wash to match. None of the Old Spice scents came close. Axe Oud Wood & Dark Vanilla antiperspirant has more vanilla and is a bit subdued but works.
Finding a body wash was harder. Cremo Spice & Black Vanilla Body Wash sounded promising, but I was apprehensive about its scent including tobacco. I did not want to smell like an ashtray. But when I opened the bottle, I was met with a surprising scent from the past. I immediately recognized it as premium Danish pipe tobacco. Most people do not recognize this aromatic scent as coming from tobacco at all. In fact, it smells more like fresh cured tobacco itself, not its smoke. This adds a very pleasant and masculine smell to the body wash on top of the spice, wood, and vanilla scent I was searching for.
The scent only lasts a few hours after you rinse off. One push on the dispenser puts enough soap on a shower loofah to wash your whole body. I do not rinse off my loofah; I squeeze the soap out of it to wash my face, then I place it back in its tray next to the bottle. The scent of the remaining soap acts like incense until my next shower.
I was also pleasantly surprised to find that this body soap has a rich lather and cleans as well as advertised. I had forgotten the reason I bought the body soap until I used it.
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Reviewed in the United States on February 27, 2026
★★★★★ 5
Intoxicating Smell!
My 'old' shower gel was getting dangerously low prompting a search for a replacement. I looked over recommendations of various sources including Amazon which led me to this Cremo product.
Over the years I've purchased just a handful of products that were either colognes, soaps of various types, or aftershaves, which really stood out for their intoxicating scent. Additionally, they might have had the incredible ability to have a lasting effect of several hours. These products stood out in a way that they also were noticed by the people around me, commenting on the great odor and inquiring as to what it was.
This Cremo product is in that category. The first time I used it clearly put it in that rare product list. Additionally, some of the other qualities of this product that others have commented on I have found to be true. A little goes a long way! You don't need to use nearly as much as some other products that I've had in order to get it to effectively wash body or hair. It truly does leave a soft feeling after using. I didn't realize how big the container would be when I purchased it, however can now say after a few weeks of using it, this is going to last me a long time! And I'm definitely pleased about that. I guess you can say that there are definitely other less-expensive products out there, but when you consider how well this product performs, it's a great buy!
If you're currently reading this, examining products for yourself, wondering if this one is for you....hesitate no longer. When this container runs out....it's definitely going to be a long while....I will definitely buy it again.
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Reviewed in the United States on January 17, 2026
★★★★★ 5
Warm base of vanilla, with a bit of spice to keep things interesting! Definitely a keeper!
First, I need to admit that Cremo is one of my “go to” brands for body wash. I love the fact that the scents are high-quality without getting into the weird “axe” arena of being able to be smelled a mile away.
The body wash has a fresh, warm, earthy smell to it. Vanilla base with a bit of spice to not make it smell like a sugar cookie. I think you all know those body washes that are just too heavy with the vanilla smell. The price for this level of body wash is spot on for what you get from a quality, moisturizing, and skin care perspective. It doesn’t dry out your skin and even the deep rich color of the gel screams quality.
This is a really great new scent from Cremo and one that I’d definitely come back to and keep in my rotation.
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Reviewed in the United States on April 25, 2026
★★★★★ 4
Good product but smell doesn't last
Cleans and rinses great, does not dry out your skin. Smell great, not as strong as the Italian Bergamot, nor does it last after showering. Once you dry off, smell pretty much gone. Bergamot lasts a little better
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Reviewed in the United States on December 19, 2025
★★★★★ 5
Beyond my expectations…
Scent: Incredible 10/10. The entire shower smells like a spa afterwards. Woody notes, vanilla scent is deliciously heavy and warm, and even one pump delivers a punch of fresh vanilla scent to your entire body. I’m a woman who is in love with heavy earthy scents and this is it!
P.S it also matches well with scents like vanilla, cocoa butter, chocolate, or even delicious “smore” scents! I pair it with vanilla and cocoa butter scented deodorant, vanilla perfume, or vanilla body lotion and it works amazing!
Longevity: Lasts the entire day if I’m staying home just relaxing, not working out or sweating very much. I could shower in the early morning and by the time my head hits the pillow I still smell it faintly!
If I’m out and about it still lasts hours I’d estimate around 6-8 hours before the scent fades! If I shower at 6am, come home at 5pm-6pm after work I still smell fresh! (I always shower 2x a day but you don’t even need to with this!
Value for Money/Size: Oh absolutely! I think I bought this soap two weeks ago? Used it nearly every day, and you can’t even tell! One pump is enough, two pumps MAX per shower per day.
I decided to lavish myself by buying this soap, because it’s a bit more pricey. But it’s actually saving me more money, at this rate I’ll be buying another one of these every 3 months! So that’s…4 times a year? 5 max? That’s incredible. I’ll probably stick with Cremo for life honestly. And I’m a woman!💕🎀
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Reviewed in the United States on March 7, 2026