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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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4.7 ★★★★★
Based on 28 reviews
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Product Reviews
★★★★★ 5
Functional and easy to install!
Color: Cool White
This umbrella light is super handy! It hinges open to allow installation around the umbrella pole, so you don’t have to remove umbrella from table to add the light. I like that it can be charged with included USB cord or can use regular batteries. The different light settings are also nice depending on how much light you need or want at the time. We’ll be using this quite often when sitting at the table by the pool this summer.
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Reviewed in the United States on July 21, 2025
★★★★★ 5
Different than most on the market worth a try!
I’ve tried a lot of pimple patches and these actually feel different from most of the ones on the market because of the sulfur/OptiMSM ingredient combination. They don’t just cover the spot — they seem to really help bring everything to the surface faster. I had one deep pimple that had been under my skin for so long I honestly started wondering if it was just a scar at that point, and this patch finally helped it come out and heal. The patches are super thin and blend into the skin really well, so they’re easy to wear during the day or overnight. They stick well without feeling harsh on my skin, and I also tried them on my tween sons, who definitely need to wash their noses better sometimes, and they seemed to work really well on their softer skin too without causing irritation. I also love the little travel pouch included. Definitely one of the more effective acne patches I’ve used.
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Reviewed in the United States on May 25, 2026
★★★★★ 4
Halo patches
I tried the Halo Dot Ultra Thin Pimple Patches, and overall I’m really impressed with them. In terms of effectiveness, they work well at helping bring pimples to a head and protecting spots overnight so they don’t get worse or irritated. The value for money is good since you get a decent amount in the pack and they’re thin enough to use during the day or night without being too noticeable. They’re super easy to use—just clean skin, place the patch, and go—and I like that they stay in place pretty well. I also find them pretty versatile, since you can wear them at home or out and about if you get a discreet one. For skin sensitivity, they’ve been gentle on my skin with no irritation, which is a big plus for me. Overall, they’re a simple, effective skincare staple I’d definitely keep on hand.
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Reviewed in the United States on June 1, 2026
★★★★★ 5
Nice patches -- Fair price -- has sulfur!
This is a review for the Halo Dot™ Pimple Patches for Face, Hydrocolloid Acne Patches with OptiMSM®, Visibly Shrinks Zit & Whitehead- Comes with Travel Essentials Inner Pouch (33 PATCHES) ($12.59)
PROS: Price -- a fair deal for 33 patches and consistent with pricing of most other similar patches online. The inclusion of sulfur (this OptiMSM business) makes these unique also -- I can't seem to find others on Amazon with similar formulation, so these are intriguing. Sulfur is known for antimicrobial properties and can be helpful for zits and a little bit gentler on your skin than benzoyl peroxide or salicylic acid -- but anytime I put any sulfur stuff on zits that are above or around my eyes, it seems to migrate INTO my eyeball while I'm asleep and man is that uncomfortable. This little patch solves that dilemma nicely by keeping the sulfur put.
CONS: none that I can see; these are a nice entry into the hydrocolloid acne patch space.
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Reviewed in the United States on May 13, 2026
★★★★★ 3
Disappointing results and poor adhesion.
I love Hydrocolloid pimple patches. I have used at least half of the brands available for sale and even the non brand ones. I’ve also used the hydrocolloid sheets and cut them to size and the hydrocolloid gel bandaids and used for pimple patches. I love how they help along the healing and keep me from picking at the pimple.
This brand I wasn’t impressed by. None of them stayed on my face for half the night. The one that remained in place the longest out of the 4, didn’t show any signs of improvement. The longest they kept in place was 3 hours. I had to reapply and I still handbrake noticed any signs of healing after using one full sheet on 4 smaller sized blemishes because the adhesive wasn’t working.
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Reviewed in the United States on May 20, 2026