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Description
Human PPIC 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 uses 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 capture antibody against Peptidyl-prolyl cis-trans isomerase C (PPIC). After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of Peptidyl-prolyl cis-trans isomerase C (PPIC) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Peptidyl-prolyl cis-trans isomerase C ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Peptidylprolyl isomerase C (PPIC), also known as CYPC or Cyclophilin C, is an enzyme encoded by the PPIC gene. As a member of the peptidyl-prolyl cis-trans isomerase (PPIase) family, this protein catalyzes the cis-trans isomerization of the proline-imidazole peptide bond, enabling it to facilitate protein folding or repair. Furthermore, it is involved in numerous biological processes, including mitochondrial metabolism, apoptosis, redox reactions, and inflammation, as well as related diseases and conditions such as ischemia-reperfusion injury, HIV/AIDS, and cancer. | |||||||||||||||||||||||||||||||||
| 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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4.3 ★★★★★
Based on 276 reviews
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Product Reviews
★★★★★ 3
Not indestructible
Size: Large, Color: Dinos Frills (Gray)
When I googled “indestructible stuffy dog toys” this popped up. My 9 month old pup loves stuffies but destroys them quickly and eats stuffing if not caught quickly. Well, this indestructible toy is cute but my dog had it torn apart in under 4 minutes so…🤷🏻♀️
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Reviewed in the United States on January 14, 2026
★★★★★ 5
Great for Tough chewers!
Size: Large, Color: Dinos Frills (Red)
My dog is a tough chewer and absolutely loves this toy! He was able to get a little stuffing out of the leg so far and it’s been about a week. This might sound like a short amount of time but it usually takes him under 15 mins to unstuff a large toy even if they are advertised for “tough chewers!” This is a very large size toy and would be good for any larger breed. He carries it around the house and constantly keeps his Dino with him. Since Ive found GoDog toys I’ve only been purchasing them because of their durability. I’ve tried Kong and Bark toys previously and they just do hold up long and are usually more expensive. This toy is a great value and dogs love them!
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Reviewed in the United States on October 20, 2025
★★★★★ 5
Super cool strong dog toy!
Size: Large, Color: Dinos Frills (Red)
Super cool strong dog toy!
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Reviewed in the United States on May 12, 2026
★★★★★ 4
LOVE this brand. Have many of their styles. This is basically an unstuffed toy.
Size: Large, Color: Dinos Frills (Gray)
I have about 10 -12 different styles of the GoDog brand toys. It is like Xmas when my Malinois dogs get a new GoDog toy. Have loved this brand for many years and have some toys which have lasted 5-6 years -- minor "surgeries" stitch they back together -- until I retired them. They have never just worn out. Just to be clear:
THESE TOYS ARE NOT CHEW PROOF. NO STUFFED TOY IS CHEW PROOF.
However, this brand of stuffed toy is more chew proof than other brands. But if you are looking for a chew proof stuffed toy be advised -- IT DOES NOT EXIST!! No fabric is chew proof. Even if they invented such a fabric I would not want to ruin my dog's teeth by letting them try to chew through it. Protecting one's dog's teeth should be the highest priority.
If a dog can chew through a PLASTIC KENNEL and even METAL WIRE KENNELS (yes, it happens) then they can easily chew through a puffy, fluffy stuffed toy. The "Chew Guard Technology" language may be misleading to some customers who are disappointed in the brand. I defend the brand and have used it for many years. No, I don't get paid or get free toys.
Since I have had many styles within this brand and even have the pink version of this style I thought I knew what I was getting. This is basically a toy without stuffing which some people prefer. For whatever reasons my dogs prefer toys that are stuffed. Really stuffed with a firm density that they can sink their teeth into. So this toy will be OK. I can tell it will not be one of their beloved toys like some of the brand's other styles such as the purple brontosaurus. My fault for not reading more closely and assuming the grey version would be exactly like the pink version which is fully stuffed. If they get bored with these two I keep some of the Chew Guard toys stock piled.
BTW one can wash and dry these toys. Yes, eventually the squeakers suck up some water and die but one can replace the squeakers. Also on Amazon. So I am willing to replace the squeakers a few times in the life time of one of these toys to be able to wash and dry them occasionally. Usually takes a couple drying cycles to get the heavily stuffed toys dried completely.
If you have an aggressive chewer you may wish to look into Goughnuts dog toys or other hard rubber toys that are designed for aggressive chewers. Even then, with a determined dog, the Goughnuts toys can be chewed through. I think they offer a replacement guarantee. The other brand which has some toys of hard rubber is West Paw. An excellent brand that also offers a one time replacement. Bully Sticks are another option for chewers.
PLEASE, PLEASE, PLEASE stay away from the hard plastic chew toys like the Nyla Bones. Hard plastic toys will destroy your dog's teeth over time. I learned the hard way (no pun intended) and ruined one of my dog's teeth. Vet asked if I allowed my dog to chew ROCKS !!!!!! Every tooth was cracked or fractured. Not wishing to pull teeth I had to pay for some very expensive root canals and caps. Vet's advise: If you can't dent the toy or chew with your finger nail DO NOT offer to your dog. Good advise to follow. Even some of the Bully Sticks (Costco Brand) are so hard they can chip or fracture teeth.
Best for you and your dog.
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Reviewed in the United States on September 1, 2019
★★★★★ 5
Well made, minor issue was quickly resolved
Size: Large, Color: Dinos Bruto (Purple)
I purchased the Dino Bruto Dog toy for my labradoodle who is quite fond of squeaky, squawky stuffed toys and oddly enough has shown a preference for purple. I opened the package with my hound standing by waiting for his gift. I detached the tags and attempted to squeak the toy. It seemed difficult to squeak or honk. I tossed the purple Dino Bruto to my dog who excitedly chomped to squeak. The toy squawked a couple times and then just quit. My poor hound was staring up at me with that "what's up with this" look on his face. I tracked down the website for the GoDog toys, not going through the Amazon return process. I called the good people at GoDog and stated simply that my dog's favorite part of the toy failed quite quickly and I was wondering about guarantees. I was told to go to the website and file a replacement request. The replacement request required that I include a photo which I did as directed. I received an reply email within a couple hours which stated that a new toy would be sent out shortly.
I am happy to report that we received a replacement toy, not the purple Dino, but a colorful wood duck. The squawker device works well and is still performing like new more than a month later.
It appears that the squeaker is not those little plastic squeakers but the bigger heavy plastic bag, bladder, with what I visualize as a duck call sort of sound device. I am guessing because I haven't seen the inside of the GoDog toys. I have seen the guts of other stuffed toys which use this type of noise device. While our dog seems to enjoy the sound of the plastic bag, bladder set-up squawkers, they seem to fail quickly and I tend to avoid them because of this. The toys using this type of squawker that I have seen, use what looks like a cheap lightweight plastic bag which is not securely attached to the "voice box".
Since the GoDog replacement toy is holding up so well, I am guessing GoDog uses a heavier grade plastic bladder coupled with reinforced construction. The purple Dino Bruto toy still looks new even after multiple times of our hound grabbing the purple toy, shaking it at us with his invite to give chase or play keep away. Since the squawker failed so quickly with the new toy, I figure it was just a defective item which does not reflect the general quality of toys from GoDog.
One evening our neighbor was visiting while his pitbull was playing in our yard with our dog. Our patio door was open so our dog ran into the house and grabbed his purple Dino Bruto toy. He is a bit of a show-off with new toys. Our dog then went racing back into the yard, shaking his GoDog toy at the pitbull. A quick game of tug-of-war ensued. Since this is a new toy, our neighbor leaped into the game and took the toy away from the dogs. In that short time of tugging between the dogs, the toy held together and did not even pull apart at any of the seams. There were no puncture marks either.
This is a game our dog has initiated before with the neighbor pitbull. Normally, the stuffed toys are ripped apart almost immediately. I was surprised and impressed with the quality of the sewing which appears to employ some sort of reinforced construction or heavier material.
I plan to make more purchases from GoDog toys. The toys are a little more expensive than some stuffed dogs toys but they hold up so well I figure they are worth a few more dollars. They are great toys that hold up well and the company provides quick responses to inquiries. I consider this a winning combination.
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Reviewed in the United States on June 15, 2013