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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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4.3 ★★★★★
Based on 18 reviews
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Product Reviews
★★★★★ 5
Soft, Lightweight, and Perfect for Year-Round Comfort
Color: Polar Blue, Size: King (90 in x 108 in)
This Berkshire 3-layer fleece blanket is exactly what I was hoping for. It is incredibly soft and has a cozy cotton-like feel without being too heavy or bulky. The king size is generous and fits the bed nicely, but it is also comfortable enough to use on the couch.
I really like that it feels warm and breathable at the same time, so it works well for different seasons. The Polar Blue color is soft and pretty, and the blanket has a nice textured look that makes it feel a little more elevated than a basic fleece blanket.
It also washes well, which is a huge plus. After washing, it still feels soft and comfortable, and it did not lose its shape. Overall, this is a great blanket if you want something cozy, lightweight, and easy to care for. I would definitely recommend it.
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Reviewed in the United States on May 10, 2026
★★★★★ 5
I love this because it folds up easily & does not take up much space but keeps you warm and secure.
Color: Southern Blues, Size: King (90 in x 108 in)
Heavy and BIG. The color is great. Its not very thick but it does have some heft to it. I love this because it folds up easily and does not take up much space but keeps you warm and secure. Its got a waffle type contour texture. It says its fleece but to me it did not feel like fleece. However, I have never heard of berkshire fleece before-- I would not guess this is fleece at all. Its not fuzzy in the least. It is breathable and the color is muted and perfect. For a king sized blanket this is surprisingly portable yet substantial enough to keep the entire bed covered and warm!
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Reviewed in the United States on May 21, 2026
★★★★★ 3
Thin, summer weight throw, quality could be better
Color: Southern Blues, Size: Throw (50 in x 60 in), Color: Southern Blues, Size: Throw (50 in x 60 in)
This is a very light weight blanket and the "fleece" is no where to be seen. The blanket looks like cotton but has a very synthetic feel to it. Three layers implies some weight but it is thin.
The construction is sloppy, see images, where the stitching looks slap dash to the point where it cannot be reversed.
I liked the color, the blue is a little on the aqua side but it is a very nice calming hue. The stitching is quilt like and nicely done.
I noticed other reviews felt like it was a medium weight but the only time I would use this would be to use as a cover if the AC went out. To me, the blanket is decorative and not particularly useful.
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Reviewed in the United States on May 15, 2026
★★★★★ 4
Average quality lightweight throw, not sold on the color
Color: Southern Blues, Size: Throw (50 in x 60 in), Color: Southern Blues, Size: Throw (50 in x 60 in)
This blanket is fairly soft and lightweight, and better suited for the summer months. I don't think the texture and "Southern Blues" color looks as nice in person as in the photos; less cozy and more like a spare blanket you might find in a hotel closet.
Interestingly the care tag says that the outer faces are cotton, but the blanket feels polyester and the content tag indicates the same. The stitching and durability feel ok, and it will probably hold up for the occasional machine washing and low heat drying.
Overall it's a fine warm season throw but not particularly notable. I would probably recommend one of the darker colors.
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Reviewed in the United States on May 23, 2026
★★★★★ 5
So easy to wash!
Color: Southern Blues, Size: Full/Queen (90 in x 90 in), Color: Southern Blues, Size: Full/Queen (90 in x 90 in)
Two things I love about this coverlet:
1. It's so much easier to wash than a duvet! Or even a duvet cover or a regular blanket, for that matter. The queen-size coverlet fit easily in my washer, and I could have put in another if I had had one. It stayed loose and balanced in the machine - the washer didn't have to readjust its balance during the spin cycles. In the dryer, it didn't wad up into a ball that had to be shaken out every few minutes. It came out feeling softer and looking good, with no wrinkles.
2. It's thinner than a duvet and suitable for warm weather, yet those three stitched layers give it substance and weight that I find more comfortable than sleeping under a pile of fluff or a bed sheet.
The soft pastel green color with cream lining (not really reversible due to the hem) is fresh and reminiscent of the 1950s coverlets that my grandmother made. Crocheting was one of her passions, and she'd create huge lacy doilies that lay across the foot of the coverlet. Very pretty look.
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Reviewed in the United States on May 1, 2026