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Description
Mouse Col6a1 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 freeze-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 freeze-thaw cycles 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 1000×g for 20 minutes, and collect the supernatant for analysis. Preparation for the Assay: 1. Remove the reagent kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare the gradient working solution of the standard: Add 1 mL of universal diluent to the lyophilized standard. Let stand for 15 minutes to completely dissolve, then gently mix (concentration 40 ng/mL). Then dilute to the following concentrations: 40 ng/mL, 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 40 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 20 ng/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 uses 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 Collagen Type VI Alpha 1 (COL6a1). After incubation and washing, the assay is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of Collagen Type VI Alpha 1 (COL6a1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse COL6a1 ELISA Kitt | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| 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 | 0.62-40 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.1 ★★★★★
Based on 20 reviews
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Product Reviews
★★★★★ 5
It is real and natural. No synthetic- no allergy!!!
Size: 2' x 6' (Double Pelt), Color: Brown Tipped
SUPER QUALITY, the color is exactly like on the picture, I love it!!!
Thank you!
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Reviewed in the United States on April 3, 2026
★★★★★ 5
Soft and luxurious, high quality - but don't buy if you have cats
Color: Ivory White, Size: 2' x 6' (Sheepskin Shape)
I give the rug a five-star rating because it deserves it. I was really surprised at how good the rug is right out of the package (it was packaged very well too). Because this rug is relatively less expensive than many other listings of natural sheepskin rugs on Amazon and elsewhere, I wasn't expecting much. But it turned out to be thick, soft, fluffy, plush, and luxurious all over. Many years ago I have purchased another natural sheepskin rug from Amazon described to be of the same size as this one (I got the 2'*6' one), but this one is definitely much bigger in its actual size. It's hands-down a high-quality, high-value purchase if you want a nice natural sheepskin rug. I also love how beautiful the natural creamy white color is.
Now, the story takes a turn. It does have a "natural and organic" smell as mentioned in many other reviews. The smell itself is not repulsive. I don't know how long it will take for it to air out (my last rug never really got rid of the smell, but it was not a big issue since I put it on the floor. I lost the rug in cross country moving.) But now I have a cat! My cat just couldn't resist tackling the rug the moment it was laid on the floor. She probably imagined herself to be a lion or a tiger who just caught some big prey. She couldn't stop kicking and biting the rug, trying to tear it apart. She came back to it a few times after breaks already. And this is day one. I am guessing this smell does stimulate some instinctive call from her genes. In a few short moments, she managed to tear off quite some fur from the edge of the rug... The rug still looks fine since I just received it today. I am not sure how long it will last with my cat though. Before ordering, I thought about this possibility and googled whether cats and sheepskin rugs can co-exist. I got many cute picture results showing angelic cats resting contently in the long and soft sheepskin fur, looking totally adorable and peaceful. Many articles say that natural rugs calm our fluffy friends down. Well, I told myself "perfect!" Win-win for me and my cat. Now, here we are, the beginning of my cat's conquest of an imaginary lamb (which was a real lamb.) I cannot even return it since it has already been damaged by my cat... In the worst case, this rug would become a really expensive cat entertainment piece that would eventually be destroyed....:((( I even started feeling guilty for buying animal-based products - but then comforted myself by remembering reading somewhere that cow and lamb leather goods come from animals that would be slaughtered for meat consumption anyways. My cat is already eating canned food containing Australian lamb, etc, etc. Really need to stop the thoughts.
So this is my review. Beautiful rug of excellent quality and unbeatable value, unless you have a cat(s).
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 3, 2022
★★★★★ 5
Good quality, nice looking, cozy
Color: Cream Brown, Size: 2 x 6 ft Sheepskin
Good quality, accurate color. Perfect length for floor beside queen size bed.
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Reviewed in the United States on May 24, 2026
★★★★★ 5
Perfect for a queen bed
Color: Tan, Size: 2' x 6' (Sheepskin Shape), Color: Tan, Size: 2' x 6' (Sheepskin Shape)
I LOVE this rug. I’m so glad I bought this instead of a cheap dupe. Reasonably priced, arrived quickly, doesn’t smell. It’s super soft and fluffy and really dense, so far I haven’t noticed any shedding. The size is perfect for draping across a queen bed. The color is beautiful and feels very natural. I might buy a second one for my bus conversion! Thank you so much!
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Reviewed in the United States on May 5, 2026
★★★★★ 5
It's perfect!
Color: Snow Grey, Size: 2' x 3' (Sheepskin Shape)
Gorgeous and soft! We love it! Very quality fur.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 20, 2026
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