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Description
Human ccr6 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 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 Chemokine CC-Motif Receptor 6 (ccr6) 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 Chemokine CC-Motif Receptor 6 (ccr6) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Chemokine C-C-Motif Receptor 6 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | CCR6 is a CCR chemokine receptor protein encoded by the CCR6 gene. It has recently been designated CD196 (cluster of differentiation 196). The gene is located on the long arm of chromosome 6 (6q27), on the Watson (plus) strand. It is 139,737 bases long and encodes a 374-amino acid protein (molecular weight 42,494 Da). This protein belongs to the A family of the G protein-coupled receptor superfamily. The gene is expressed in lymphoid and non-lymphoid tissues, such as the spleen, lymph nodes, pancreas, colon, appendix, and small intestine. It is expressed on B cells, immature dendritic cells (DCs), T cells (Th1, Th2, Th17, Tregs), natural killer T cells (NKT cells), and neutrophils. This chemokine receptor is unique because, in contrast to other chemokine receptors, it binds only one chemokine ligand, CCL20. This receptor has been shown to be important for B lineage maturation and antigen-driven B cell differentiation, and it may regulate the migration and recruitment of dendritic cells and T cells during inflammation and immune responses. This gene has been described as an alternatively spliced transcript variant encoding the same protein. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, and other biological fluids |
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4.5 ★★★★★
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Product Reviews
★★★★★ 5
Powerful and silent!
Color: Lux Metallic Blue, Size: Rechargeable Z1 Motor Max
Wow, call me impressed! I just got the rechargeable version in blue and it's gorgeous! The metal shaft is thick, high quality and long enough to reach the bottom of a tall mug. And the double whip end works like magic. I had a cheaper frother (another brand) and this one blows it away! I couldn't even get much of a froth with my other one and having to hold the button the whole time was annoying. One press of this button and my milk was frothed in seconds. I didn't even use the higher setting! My morning latte is very happy.
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Reviewed in the United States on March 4, 2026
★★★★★ 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
Strong and Quiet
Color: Lux Cardinal Red, Size: Rechargeable Z1 Motor Max
Better than I expected. Works in two modes. Makes mixing a breeze.
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Reviewed in the United States on May 23, 2026
★★★★★ 5
Powerful!
Color: Lux Galaxy, Size: Rechargeable Z1 Motor Max
This thing is powerful and works great! I was hesitant about some of the negative reviews, but this is worth it so far. I use it every day and haven't charged it in the week that I've had it. Some people complained about turning it off.... It's not that hard. Just hold the button down for a second or two and it shuts off, no fuss. If you tap/press the button you'll cycle through the next speed and then off. I only use the first speed to make my protein shake and matcha. I feel like the high speed would just whip everything to pure foam! I'm hoping this lasts me a long time.
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Reviewed in the United States on May 5, 2026