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Description
Mouse CK18-M30 ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. 2. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. 3. 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 the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL 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 the supernatant can be assayed. 4. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend 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 freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. 5. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 6. 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 cytokeratin 18-M30 (CK18-M30) capture antibody. 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 cytokeratin 18-M30 (CK18-M30) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Cytokeratin 18-M30 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Cytokeratin (CK) is the largest and most complex subfamily of cytoskeletal intermediate filament proteins, of which CK18 is a member. It primarily consists of two fragments, M30 and M65. The M65 fragment in plasma reflects the total amount of CK18 released into the blood following cell necrosis, while the M30 fragment has become a clinical marker of apoptosis. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.6 ★★★★★
Based on 1003 reviews
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Product Reviews
★★★★★ 5
Good for small dogs
Size: Small - 2 Count (Pack of 1)
These are great for small dogs that have trouble getting their mouth around a regular size tennis ball. My only issue is that they are small enough to roll under things and I end up having to move furniture to find them. My dog loves them.
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Reviewed in the United States on February 13, 2026
★★★★★ 5
Chuckit! Max Glow Ball Dog Toy - A Canine Delight in the Dark!
Size: Small, Pattern Name: Pack of 1
I recently introduced the Chuckit! Max Glow Ball Dog Toy to my furry friend, and it has quickly become a favorite in our playtime routine. Here's why it deserves a glowing 5-star review:
**Pros:**
**1. Day and Night Fun:**
The Max Glow feature of this ball adds an extra layer of excitement to playtime. Whether it's a sunny day or a moonlit night, my dog can enjoy fetching and playing with this ball anytime, making it a versatile and engaging toy.
**2. Long-Lasting Glow:**
The glow effect lasts surprisingly long, providing extended play sessions even after the sun sets. The consistent glow adds a thrilling element to our nighttime activities, making it a standout feature of this toy.
**3. Durable Design:**
The Chuckit! Max Glow Ball is built to last. Its durable construction can withstand the enthusiastic chewing and fetching antics of my dog, ensuring longevity and making it a reliable addition to our collection of toys.
**4. Easy to Clean:**
Cleaning this ball is a breeze. Whether it's muddy paw prints or slobber, a quick rinse under the tap is all it takes to keep the Chuckit! Max Glow Ball looking and feeling fresh for the next round of play.
**5. Compatible with Chuckit! Launchers:**
The ball is compatible with Chuckit! launchers, adding an extra layer of fun to our play sessions. The launcher allows for longer throws, providing a good workout for my dog and adding an interactive aspect to our outdoor activities.
**6. Fetching Favorite:**
My dog absolutely loves fetching this ball. The texture and size make it easy for him to carry in his mouth, and the glowing feature captures his attention and keeps him engaged during playtime.
**7. Versatile Outdoor Toy:**
Whether it's a game of fetch in the backyard, a trip to the park, or an evening stroll, the Chuckit! Max Glow Ball has become our go-to outdoor toy. Its versatility and engaging qualities make it an essential part of our canine adventures.
In conclusion, the Chuckit! Max Glow Ball Dog Toy has brought endless joy and excitement to our playtime routine. Its durable design, long-lasting glow, and compatibility with Chuckit! launchers make it a 5-star favorite in my dog's toy collection. If you're looking to add a touch of magic to your dog's play sessions, this glowing ball is a must-have. Let the nighttime playtime adventures begin! 🌙🐾🌟
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Reviewed in the United States on January 26, 2024
★★★★★ 5
I've bought like 20! Love them!!!
Size: Large, Pattern Name: Pack of 1
My dog is obsessed with this ball! Glow in the Dark is super bouncy, easy to spot at night, and tough enough to handle rough play. Perfect for evening fetch sessions—fun for both of us! We have bought about 20 of these so far. I always bring my dog to play fetch at night, and this is perfect if you're playing in a field or beach. Glows in the dark so both you and your dog can see it. The size of the ball is great for my big dog. It is also the perfect type of silicone so that my dog cannot easily tear it apart and chew it, which is GREAT! We do NOT want our dog's ingesting silicone at all. If you have a ball chewer, this ball is perfect and durable. Chewing ability is good and is not too tough but tough enough.
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Reviewed in the United States on August 10, 2025
★★★★★ 5
Cane Corso approved....
Size: Large, Pattern Name: Pack of 1
My Cane Corso loves this ball!!! She carries it around in her mouth like a round pacifier.
Sometimes I'll wake up with the ball next to my head on my pillow. Lol
This is the third one I've bought.
She doesn't destroy them at all . Somehow she loses them. Yesterday she had two floating around. Lol.
Earlier I gave her one of those hard rubber balls with feet... She destroyed it within minutes. She can skin a tennis ball and chew it in half in about 5 minutes.
This ball is by far a favorite toy of her's. And she doesn't chew it to bits .
And it glows in the dark which is fun for playing fetch at night.
It's very easy to clean.
Sometimes I put little snacks inside for her while she's laying down with her ball.
I will definitely keep buying these balls everytime we lose one . They make her happy. And that makes me happy.
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Reviewed in the United States on May 12, 2026
★★★★★ 5
BEST GLOW BALL! MY COLLIE BLUE HEELER MIX IS OBSESSED
Size: Large, Pattern Name: Pack of 1
Well if I could just say...Mommas a fan too....BUT I think my dog should be the sponsor of CHUCKIT because he owns everything and he BREATHES, DREAMS, RUNS with #chuckit balls!! Since 8wks. old & Now 10 yrs young, BDAY=8.10.26....he wakes up with it in his mouth and falls asleep with IT! I HAVE THEEEE PROOF LOL SERIOUSLY OBSESSED! #chuckit #makemydogyoursponsor WE HAVE EVERY BALL X10 FOR SOME, launchers, bucket o balls , you name it so yeah #obssesedisanunderstatement #bordercollieblueheelermix #HARLEY ♡ nice colors and the durability is amazing 👏 know if you have a chewer I'm not sure because he just likes to catch the ball and then chew it sometimes and then leave it alone, until next time around. But the GLOW BALL just being cewed for so long and the textire is obviously different so it does end up cracking and you throw it away, definitely different rubber but highly recommend them both actually ALL of them👌🎉🥳🫶🏼 my Sisters 2 yr old pit destroyed them in minutes but only FOR PLAY , so IF YOU DONT WANT THAT BALL chewed to pcs. Put it up LOL and oh
all of the balls EXCEPT the GLOW BALL stay true to form and durability, for real Harleys had some of them for years!! And nothing wrong with the CHUCKIT ball 10☆☆☆☆☆☆☆☆☆☆ SIZE is perfect too he love the medium 2.5" ball. Perfect for catching for hrs on end. You also have a choice you can get squeaky, crunchy,or nothing or glow in the dark, 3 sizes to fit your pupper..
.. so many choices and it doesn't have to be a ball you can pick from a ton of toys ' tuggers ,launchers and indoor toys too. As for the Chew resistance...mm touchy...like I said. Aggressive chewer are more likely to tear it up way quicker than the just playing mainly like I want to fetch dog. Either way. I dont purchase any other brand besides chuck it. Has always my pick, like 20+ yrs now lol. Hands down. This is not a paid thing or promoted or whatever. Just felt i should review for chuck it♡ to date has made my dog happy and healthier for years to come♡ thanks
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Reviewed in the United States on May 8, 2026