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Description
Mouse CC motif 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: 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: 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. 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. 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. 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. 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. 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 a capture antibody against Chemokine (CC motif) ligand 6 (CCL6). 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 Chemokine (CC motif) ligand 6 (CCL6) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Chemokine ligand 6 (CCL6) ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Chemokine CC-motif ligand 6 (CCL6) is a small cytokine belonging to the CC chemokine family. It is expressed in cells of the neutrophil and macrophage lineages and can be strongly induced under conditions conducive for myeloid cell differentiation. It is highly expressed in bone marrow cultures stimulated with the cytokine GM-CSF. Lower levels of gene expression are also observed in certain myeloid-derived cell lines (such as the immature myeloid cell lines DA3 and 32D cl3, and the macrophage line P388D), which are also strongly induced when cultured with GM-CSF. However, CCL6 expression is greatly reduced in activated T cell lines. It can also be induced in the lung by the cytokine interleukin-13. The cell surface receptor for CCL6 is believed to be the chemokine receptor CCR1. | |||||||||||||||||||||||||||||||||
| 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 | 0.15-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.8 ★★★★★
Based on 112 reviews
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Product Reviews
★★★★★ 5
Powerful, rechargeable battery, easy to use & clean
Color: Lux Absolute Silver, Size: Rechargeable Z1 Motor Max
2 speeds, easy to change back and forth to different speeds with one click , easily blends 1/4 cup protein powder in 12 oz milk on low speed. Easy to clean: Stick blending wand in tall glass with warm water + dish soap, turn on low speed to clean easily (don’t stick the part that holds the battery in glass). Also of note, chosen by Americas Test Kitchen as best milk frother but I use it to make protein shakes.
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Reviewed in the United States on June 5, 2026
★★★★★ 5
Works great on bullet coffee!!
Color: Lux Alpha Black, Size: Rechargeable Z1 Motor Max
I am really pleased with this frother! Frothers I have used in the past didn’t mix my decaf coffee, 2% milk, and MCT oil as thoroughly as I like (oil and coffee separated). This frother blends everything AND I get a nice layer of frothy milk on top!!! Charge lasts a long and the froth can be submerged all the way up to the button!!!
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Reviewed in the United States on June 5, 2026
★★★★★ 5
Still The Best Frother Ever!
Color: Black
I love how it is easy to use, how it holds a charge, and how much power it has. I've only charged it once since I got it & I've been using it on a daily basis!!! Though it isn't a long time, I'm happy with it so far! I definitely will recommend it to anyone looking for a great frother. If you are looking for one, look no further! You won't be disappointed! I've read some comments where they found it to be too strong that their drink spilled everywhere. But for me I have not had any incident of spilling over. I have used it both for my coffee and my matcha tea and it worked great without any spilling over. You just have to start with the lowest power and ensure that you have a large enough container to have a room for your frothing. My sister tried to use it recently during her visit to make us matcha tea. However, she experienced spillage all over the cup, because she had used a small cup. Otherwise, she liked it very much and said she will get one when her battery operated one stops working...
Another thing I like about the product was the customer service that I had received. When I got my first order which came quickly, it wasn't working after having charged it over 24 hours. It was not turning on at all. I returned it and ordered a replacement which I received within a day or two. This time after charging it for the recommended amount of time, I was a bit skeptical when I turned it on. However, I was pleasantly surprised that it worked like it was supposed to and it has been working ever since having charged it only once so far. I may come back and give an update depending on how it does after 6 months of use, so keep in tune...
I don't know if there is a way to write a new entry as a follow up to the old entry, so I'm going to add my follow up comment over here.
May 23, 2025 - I love my frother more than I can say in words. It's functionality has not changed a bit from the time I have purchased it. It charge lasts several weeks to few months before I charge it again and the power is still the same. Recently I decided to buy another one for my office use. I like the white color and so tried it but that one wouldn't even turn on. And then I got a gray one to have a different color. That one was much weaker than the black one I own. Then I replaced it with another gray one within few days. I tried it and it was exactly the same as the one I already replaced. So then I decided to go with a black one again hoping that it would be the same as the one I have. The black one did much better than the gray ones in the strength or power of the frother. However, it is weaker than the one I already have been using since last August. While I am not very happy with the quality being inconsistent in all of the ones I have tried including the recent pictures of the black one, I have decided to keep the black one because it is relatively better than the gray ones I tried. What I don't understand is why there is inconsistency in the quality of the same product by the same maker. I am beginning to wonder if the black one I purchased in Augusta last year was anomaly of a quality and a good way. Maybe I lucked out on my older black frother, but I'm unsure if quality is going to get better if I were to order another one. I have a sister who is a pro in making coffee drinks and she was the reason I had ordered this frother to begin with. She uses a battery operated one at this time and I have been thinking about getting her one like mine. While I am still skeptical, I am going to give it a shot and order her one sometime soon. If it doesn't work, I guess we will return it back. But one thing that I am grateful is that I have been able to return every one of those that I have tried recently with no issues. I hope that I don't have to return my most recent purchase in black. I was not even able to review the new purchase because of the item being the same color which is crazy.
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Reviewed in the United States on September 8, 2024
★★★★★ 5
Outstanding Frother!
Color: Space Gray
Exceptional! This frother is in another league compared to the cheapo push button frothers. The motor is smooth, quiet and powerful. This has an intuitive thumb dial on top that operates smoothly with a wide range of smooth speed adjustment. It’s gentle enough to froth a little bit of milk for a macchiato and powerful enough to scramble 3 eggs. The head is removable for easy cleaning. It’s also rechargeable. I charged this the day I bought it on April first. I use it at least twice a day. It’s September 26th today and I have yet to recharge it and it’s not showing any signs of needing a charge. One minor quibble is that they give you 2 of the same frother heads. They are built so well, that I don’t imagine it ever wearing out. It would have been nice to get a whisk or something different. Not a deal breaker at all though.
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Reviewed in the United States on September 26, 2025
★★★★★ 5
Solid unit
Color: Black
Handheld Mixer/Frother Review
A Premium Choice That Transforms Morning Coffee
I am thoroughly impressed with this handheld mixer and frother. Each morning, I blend cacao and collagen powder into my coffee, and in the past, I always ended up with stubborn lumps using only a spoon or whisk. I had previously purchased a few inexpensive handheld mixers, but they disappointed me both in price and performance, struggling to mix the powders thoroughly—especially at the bottom of the cup. I came close to abandoning handheld mixers altogether, but decided to give it one last try.
After reading countless reviews and carefully comparing models, I opted to invest in a higher-end handheld mixer. The anticipation was high—would this mixer truly distinguish itself from the budget options cluttering my kitchen drawer? From the moment I unboxed it, the difference was clear. The mixer felt substantial in my hand, and the build quality reflected thoughtful engineering.
While this model cost roughly twice as much as the cheaper alternatives, it proved to be well worth the investment. Initially, I was concerned about the dial at the top, fearing it might be difficult to control. However, I was pleasantly surprised by its intuitive operation. The unit offers a solid feel, plenty of power, and a versatile range of speeds—from a gentle stir perfect for coffee to an impressively fast setting.
I had not previously considered using this mixer as a frother, but as a cappuccino enthusiast, I decided to give it a try. I heated both 2% milk and oat milk to take the chill off, then submerged the mixing wand. Allowing the mixer to rise above the milk introduced air and created a rich, creamy foam—an unbelievable result. My homemade cappuccino now tastes like something from a café.
The controls are robust, the construction is reliable, and the power is consistent. Cleanup is effortless, requiring only a quick rinse under the tap. The battery life is also noteworthy, lasting through several uses without losing strength.
If you are hesitant to spend a bit more on a quality handheld mixer, I wholeheartedly recommend making the investment. This mixer has become an essential part of my morning routine. Excellent product—well done!
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Reviewed in the United States on August 13, 2025