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Description
Human Cath-C 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 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 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 Cathepsin C (Cath-C) 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 Cathepsin C (Cath-C) 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 Cathepsin C ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Cathepsin C, also known as dipeptidyl peptidase I, is a lysosomal cysteine exoprotease that belongs to the peptidase C1 subfamily of cysteine cathepsins. It is encoded by the CTSC gene. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.1 ★★★★★
Based on 357 reviews
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Product Reviews
★★★★★ 5
Nice toy for Chewers!
Was chewed in a day or so, but she loved it.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 13, 2026
★★★★★ 5
Still in one piece and played with!
Color: Checkers - Dinos Bruto (Purple), Size: Mini
Very durable for my little King Charles Chewer. She's had it for a while now and all though the chewing phase has calmed down, she still can be somewhat rough with the toys. The little dino is standing firm. It was a battle she figured she couldn't win with Dino. Now he's a fetch/ tug o war toy that still gets a few nibbles.
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Reviewed in the United States on June 10, 2026
★★★★★ 4
DONT BUY THE ELEPHANT, ITS NO GOOD. but GR8 BRAND!
Color: Checkers - Elephant (Gray), Size: Large
The elephant toy had no trunk by the FIRST day! The elephant now looks like an angry Koala bear OR an angry trunk less elephant(whichever u want to say) I LOVE the donkey and dinosaur toys so I bought the elephant confidently, thinking I had nothing to worry about. I wouldn't have spent money on this knowing the trunk would be chewed off in a matter of minutes. My Bella usually chews but she has had her donkey and dinosaurs with the chew guard for MONTHS and there are NO TEARS OR RIPS even with her playing and chewing on them ROUGHLY all day. If you buy any of these type toys I DO highly recommend the donkey or dinosaurs ONLY. The elephant being chewed up in a matter of minutes really makes me angry that I ever spent the money on it. HAD I known it wouldn't be the high quality dog toy I'm used to from GoDog, I would've just bought a second donkey or dino! Now Bella doesn't get her new toy for the month (I try 2 buy her a new stuffed toy or 2 about 1 time a month whenever I have the extra money for it) I guess she will have to go without this month because I spent it on this now trunkless elephant/angry koala:( Whichever name you call it, it is now going to be trash.
UPDATE!
If you contact the GODOG company they will send one replacement toy for your pet! They also asked me my pet's size, breed and age in order to send the best possible toy! They sent Bella a snake that has held up Extremely WELL! It's great and so far is my favorite(and one of Bella's favorites) The snake has less sqeaky things in it so she doesn't have the drsire to tear it apart immediately to retrieve them by rippong it to shreds:) My updated rating is for the MANUFACTURER'S GREAT CUSTOMER SERVICE. The fact that they actually replaced it was good but the fact that they went that extra mile by researching the best toy match for my Bella was above and beyond! If you have a Godog toy that went bust right away, contact the manufacturer for a replacement. If you give them your dog's age, weight and breed they will even try to match your pets replacement toy with something that is better suited to your dog's chewing style. (Bella's style is destructive haha)
Good luck, ya'll!
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Reviewed in the United States on October 26, 2016
★★★★★ 5
My Dobermans favorite toy to destroy, and one of our recommended picks.
Color: Checkers - Elephant (Gray), Size: Large
Ok, here's an honest breakdown review. I have a 6 month old Doberman who shreds every stuffed animal and dog bed he's come across. He's one of the worst chewers I've owned. We have tried a WIDE variety of stuffed toys.
That said, this elephant is his favorite toy. I have ordered 4 of them so far, and just ordered another 4-pack on a buy three get one free deal. The elephant stands out above all the other stuffed creatures from goDog for me as to durability. Hence why I keep buying more. But they usually last about 2-3 weeks on average.
In our house, the poor creatures are subjected to vicious, crazy, shaking of the head, helicopter type rotation. It' so hard if it hits your bare arm or leg it actually stings especially when its wet. Then the old, put it between his legs laying down, tearing at the legs action. The limbs are usually the first things to go over time. But they are each almost self sealing, so you clean up as little fluff and keep using it. Eventually the pup reaches the main body and finishes it off.
For a normal light to normal chewer, one of two will get you a long way. For those of us resigned to having zoomie type manic chewers that need a constant supply of toys, 2-3 week durability is pretty sound too. I do not recommend the green cheaper monster. It lasted about 45 minutes. The free replacement took about 3 days. Hopefully this aids you in your decision!
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Reviewed in the United States on June 10, 2026
★★★★★ 5
Very durable for rowdy pups
Color: Checkers - Elephant (Gray), Size: Large
This toy has outlasted any other dog toy I have ever bought. I have one large and one medium dog who share toys, fight over toys, chew them to death and still play with them. It might be because it isn't one of their favorites. I don't know. All I know is that our house is littered with dog toys in different states of destruction and this elephant toy has very little damage. It has definitely been in the dog toy rotation.
I had this toy for 2 months without any major damage. The day I wrote the review the dogs tore the leg off! I still think it is a good toy. Some toys lose the stuffing or squeaker the first day.
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Reviewed in the United States on May 18, 2025